LOCUS Exported 6088 bp ds-DNA circular SYN 16-JUN-2022 DEFINITION ddGFP A-NES. ACCESSION . VERSION . KEYWORDS GA-NES SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6088) AUTHORS Ding Y, Li J, Enterina JR, Shen Y, Zhang I, Tewson PH, Mo GC, Zhang J, Quinn AM, Hughes TE, Maysinger D, Alford SC, Zhang Y, Campbell RE TITLE Ratiometric biosensors based on dimerization-dependent fluorescent protein exchange. JOURNAL Nat Methods. 2015 Jan 26. doi: 10.1038/nmeth.3261. PUBMED 25622108 REFERENCE 2 (bases 1 to 6088) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 16, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6088 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(8..28) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 13..342 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 193..328 /label=SV40 ori /note="SV40 origin of replication" primer_bind 255..274 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 409..1203 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(463..482) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1073..1092 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 1377..1498 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1414..1433) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1468..1487 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(1547..1563) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1547..1563) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1560..1582) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1571..1587 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(1595..1625) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1640..1661 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1778..1795) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1949..2537) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2029..2048) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2708..3568) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 3331..3350 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3569..3673) /gene="bla" /label=AmpR promoter primer_bind complement(3748..3767) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 3939..4318 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4319..4522 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4473..4493 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 4567..4586 /label=T7 /note="T7 promoter, forward primer" promoter 4567..4585 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 4610..4619 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(4761..4779) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 5029..5048 /label=mCherry-F /note="mCherry, forward primer" primer_bind 5223..5246 /label=DsRed1-C /note="DsRed1, forward primer" primer_bind complement(5383..5400) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5389..5613 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5659..6087 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5746..5765) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5956..5977 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 61 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 121 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc 181 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 241 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag 301 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt 361 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa 421 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg 481 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc 541 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca 601 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc 661 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca 721 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat 781 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca 841 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg 901 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc 961 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct 1021 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct 1081 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac 1141 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc 1201 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag 1261 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg 1321 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact 1381 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 1441 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 1501 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc 1561 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt 1621 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc 1681 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg 1741 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct 1801 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 1861 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 1921 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 1981 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 2041 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 2101 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctataggt 2161 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 2221 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 2281 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 2341 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 2401 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 2461 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 2521 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 2581 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 2641 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 2701 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 2761 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 2821 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 2881 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 2941 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 3001 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 3061 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 3121 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 3181 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 3241 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 3301 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 3361 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 3421 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 3481 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 3541 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 3601 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 3661 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc 3721 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca 3781 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc 3841 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt 3901 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta 3961 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac 4021 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc 4081 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt 4141 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac 4201 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac 4261 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt 4321 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc 4381 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt 4441 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg 4501 ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt actggcttat 4561 cgaaattaat acgactcact atagggagac ccaagctggc tagcgtgccg ccaccatggc 4621 ctcgagaatg gtgagcaaga gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg 4681 cttggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc 4741 ttacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc 4801 ctgggacatc ctgtcccccc tgatcatgta cggctccaag atgtacgtga agcaccccgc 4861 cgacgtcccc gattacatga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat 4921 gcacttcgag gacggcggtc tggtgaccgc gacacaggac tcctccctgc aggacggcac 4981 gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc ccgtaatgca 5041 gaagaagacc ttgggctggg attatgccac cgagcgcctg taccccgaag aaggcgtgct 5101 gaagggcgag ctcctggggc gcctgaagct gaaggacggc ggcctcaacc tggtggagtc 5161 caagaccatc tacatggcca agaagcccgt gcaactgccc ggctactact tcgtggacac 5221 caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgctc 5281 cgagggccgc caccacctgg ggcatggcac cggcagcacc ttagccttga aattagcagg 5341 tcttgatatc gggagctaaa agcttggtaa accgctgatc agcctcgact gtgccttcta 5401 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca 5461 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc 5521 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata 5581 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg 5641 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 5701 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 5761 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 5821 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg 5881 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 5941 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 6001 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 6061 tgatttaaca aaaatttaac gcgaatta //