LOCUS Exported 5103 bp ds-DNA circular SYN 04-JUN-2020 DEFINITION . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5103) AUTHORS rvieira TITLE Direct Submission JOURNAL Exported Thursday, Jun 4, 2020 from SnapGene 5.1.3 https://www.snapgene.com FEATURES Location/Qualifiers source 1..5103 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" CDS complement(1..859) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments: 1: 1 .. 790 / #ccffcc 2: 791 .. 859 / #ccffcc / signal sequence Cleavage site after base 790" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 622..641 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" /note="color: black; sequence: ATAATACCGCGCCACATAGC" promoter complement(860..951) /gene="bla" /label=AmpR promoter /note="color: #ffffff; direction: LEFT" primer_bind 993..1012 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" /note="color: black; sequence: ATCGCGAGCCCATTTATACC" primer_bind 1018..1037 /label=pENTR-R /note="pENTR vectors, reverse primer" /note="color: black; sequence: ATGGCTCATAACACCCCTTG" misc_feature 1251..1276 /label=MCS /note="color: #333333" protein_bind 1293..1392 /label=attP /bound_moiety="phage phi-C31 integrase" /note="attachment site of phage phi-C31" /note="color: #c0c0c0" misc_feature 1403..1436 /label=FRT 14 /note="FRT14" /note="color: #ff9900; direction: RIGHT" misc_feature 1480..1513 /label=FRT /note="FRT" /note="color: #ffff99; direction: RIGHT" misc_feature 1612..1642 /label=SD /note="Predicted by www.fruitfly.org" /note="color: #00ccff" CDS complement(1659..2378) /codon_start=1 /product="GFP" /label=GFP /note="color: #00ff00" /translation="GSVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLT LKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKD DGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGI KANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVL LEFVTAAGITLGMDELYK" misc_feature 2400..2439 /label=SA /note="predicted using www.fruitfly.org" /note="color: #00ccff" misc_feature complement(2519..2552) /label=FRT 14 /note="FRT 14 " /note="color: #ff9900; direction: LEFT" misc_feature complement(2603..2636) /label=FRT /note="FRT" /note="color: #ffff99; direction: LEFT" protein_bind complement(2721..2754) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." /note="color: #31849b; direction: LEFT" promoter 2818..2985 /label=3XP3 /note="color: #ffffff; direction: RIGHT" CDS 2986..3663 /codon_start=1 /product="rapidly maturing tetrameric variant of DsRedfluorescent protein (Bevis and Glick, 2002)" /label=DsRed-Express /note="mammalian codon-optimized" /note="color: #ff0000" /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH LFL" primer_bind complement(3166..3186) /label=DsRed1-N /note="DsRed1, reverse primer" /note="color: black; sequence: GTACTGGAACTGGGGGGACAG" primer_bind 3578..3601 /label=DsRed1-C /note="DsRed1, forward primer" /note="color: black; sequence: AGCTGGACATCACCTCCCACAACG" polyA_signal 3784..3905 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" /note="color: #808080" primer_bind complement(3821..3840) /label=SV40pA-R /note="SV40 polyA, reverse primer" /note="color: black; sequence: GAAATTTGTGATGCTATTGC" primer_bind 3875..3894 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" /note="color: black; sequence: GTGGTTTGTCCAAACTCATC" protein_bind complement(3912..3945) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." /note="color: #31849b; direction: LEFT" protein_bind 3965..4064 /label=attP /bound_moiety="phage phi-C31 integrase" /note="attachment site of phage phi-C31" /note="color: #c0c0c0" misc_feature 4076..4105 /label=MCS /note="color: #333333" primer_bind complement(4104..4121) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21)Forward" /note="color: black; sequence: TGTAAAACGACGGCCAGT" primer_bind complement(4219..4236) /label=L4440 /note="L4440 vector, forward primer" /note="color: black; sequence: AGCGAGTCAGTGAGCGAG" rep_origin complement(4390..4978) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" primer_bind complement(4470..4489) /label=pBR322ori-F /note="pBR322 origin, forward primer" /note="color: black; sequence: GGGAAACGCCTGGTATCTTT" ORIGIN 1 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 61 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 121 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc 181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 301 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 481 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 841 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt 901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca 961 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg 1021 gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg 1081 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc ccatgcgaga 1141 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg 1201 cccgggctaa ttatggggtg tcgcccttcg ctgaagcagg tggaattctt gcatgctcct 1261 aggtatactg cagttgctat ctattacaaa gtcccaggtc agaagcggtt ttcgggagta 1321 gtgccccaac tggggtaacc tttgagttct ctcagttggg ggcgtagggt cgccgacatg 1381 acacaagggg ttgaagactt ctgaagttcc tattctatca gaagtatagg aacttcagcg 1441 gctggattgt agctgctatt agcaatatga aacctcttag aagttcctat tctctagaaa 1501 gtataggaac ttcgaattct ggcgacttgt cgcatctttc tctgagtggg taaggtcatt 1561 gtggggtctt aagggctaaa gggtagtgat gatgaaggca tttgatgcca ttgttcaata 1621 acttacctgc cggaacctcc gcttccaccg ctacctccct tgtacagctc atccatgccc 1681 agggtgatgc cggcggcggt cacgaactcc agcagcacca tgtgatcgcg cttctcgttg 1741 gggtccttgg acagcacgct ctgggtgctc aggtagtggt tatcgggcag cagcactggg 1801 ccgtcgccga tgggggtgtt ctgctggtag tgatcggcca gctgcacgga gccatcctcc 1861 acattgtggc ggatcttgaa gttggccttg atgccgttct tctgcttatc ggcggtgatg 1921 tacacgttgt ggctgttgaa gttgtactcc agcttgtggc ccaggatgtt gccatcctcc 1981 ttgaaatcga tgcccttcag ctcgatgcgg ttcaccaggg tatcgccctc gaacttcacc 2041 tcggcgcggg tcttgtaggt gccgtcatcc ttgaagctga tggtgcgctc ctgcacgtag 2101 ccctcgggca tggcgctctt gaagaaatcg tgctgcttca tgtgatcggg gtagcggctg 2161 aagcactgca cgccgtaggt cagggtggtc accagggtgg gccagggcac gggcagcttg 2221 ccggtggtgc agatgaactt cagggtcagc ttgccgttgg tggcgtcgcc ctcgccctcg 2281 ccgcgcacgc tgaacttgtg gccgttcacg tcgccatcca gctccaccag gatgggcacc 2341 acgccggtga acagctcctc gcccttggac acggatccgg atccgccgct acctccgctt 2401 ccaccggaac ctcccgacct gcggaagaga gataaatcgg ttgattaaaa ttagttcaag 2461 aataattagt atatttaaac atgccgggga tgggacaagt cgccatgttg gatcgactga 2521 agttcctata cttctgatag aataggaact tcttgcctta acccagaaat tatcactgtt 2581 attctttaga atggtgcaaa gagaagttcc tatactttct agagaatagg aacttcccgt 2641 cttcccgcgg acatatgcac acctgcgatc gtagtgcccc aactggggta acctttgagt 2701 tctctcagtt gggggcgtag ataacttcgt ataatgtatg ctatacgaag ttatcgtacg 2761 ggatctaatt caattagaga ctaattcaat tagagctaat tcaattagga tccaagctta 2821 tcgatttcga accctcgacc gccggagtat aaatagaggc gcttcgtcta cggagcgaca 2881 attcaattca aacaagcaaa gtgaacacgt cgctaagcga aagctaagca aataaacaag 2941 cgcagctgaa caagctaaac aatcggctcg aagccggtcg ccaccatggc ctcctccgag 3001 gacgtcatca aggagttcat gcgcttcaag gtgcgcatgg agggctccgt gaacggccac 3061 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag 3121 ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc 3181 cagtacggct ccaaggtgta cgtgaagcac cccgccgaca tccccgacta caagaagctg 3241 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg 3301 accgtgaccc aggactcctc cctccaggac ggctccttca tctacaaggt gaagttcatc 3361 ggcgtgaact tcccctccga cggccccgta atgcagaaga agactatggg ctgggaggcg 3421 tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca caaggccctg 3481 aagctgaagg acggcggcca ctacctggtg gagttcaagt ccatctacat ggccaagaag 3541 cccgtgcagc tgcccggcta ctactacgtg gactccaagc tggacatcac ctcccacaac 3601 gaggactaca ccatcgtgga gcagtacgag cgcgccgagg gccgccacca cctgttcctg 3661 taggggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct 3721 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt 3781 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 3841 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 3901 tcttaaccgg tataacttcg tataatgtat gctatacgaa gttatagaag agcactccgt 3961 cttcaacccc ttgtgtcatg tcggcgaccc tacgccccca actgagagaa ctcaaaggtt 4021 accccagttg gggcactact cccgaaaacc gcttctgacc tgggaagctt gttccatccc 4081 actcgagtac gtaggtacct cagactggcc gtcgttttac aacacagaaa gagtttgtag 4141 aaacgcaaaa aggccatccg tcaggggcct tctgcttagt ttgatgcctg gcagttccct 4201 actctcgcct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 4261 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 4321 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 4381 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 4441 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 4501 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 4561 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 4621 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 4681 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 4741 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 4801 gtggtgggct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 4861 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 4921 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 4981 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgc gcgtaactca 5041 cgttaaggga ttttggtcat gagcttgcgc cgtcccgtca agtcagcgta atgctctgct 5101 ttt //