LOCUS Exported 8844 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Constitutively expresses yellow fluorescent protein , EYFP. ACCESSION . VERSION . KEYWORDS pExp_CAG:EYFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8844) AUTHORS Davidsohn N, Beal J, Kiani S, Adler A, Yaman F, Li Y, Xie Z, Weiss R TITLE Accurate Predictions of Genetic Circuit Behavior from Part Characterization and Modular Composition. JOURNAL ACS Synth Biol. 2014 Nov 17. PUBMED 25369267 REFERENCE 2 (bases 1 to 8844) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8844 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(51..68) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(222..810) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(302..321) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(981..1841) /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 1604..1623 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1842..1946) /gene="bla" /label=AmpR promoter rep_origin complement(1972..2427) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2109..2130) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2321..2340 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2569..2585 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 2595..2613 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 2628..3431 /label=HA-L /note="left homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene" misc_feature 3510..3535 /label=SA /note="splice acceptor site" protein_bind 4207..4227 /gene="mutant version of attB" /label=attB4 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" primer_bind 4309..4325 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 4332..4350 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" enhancer 4460..4763 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4765..5040 /label=chicken beta-actin promoter primer_bind 5978..6000 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" protein_bind 6112..6136 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 6150..6159 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 6156..6875 /codon_start=1 /product="enhanced YFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(6201..6222) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(6462..6481) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 6809..6830 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(6877..6901) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(6991..7010) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 7056..7111 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(7110..7129) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(7472..7488) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 7496..7512 /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(7520..7550) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7565..7586 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." misc_feature 7759..8595 /label=HA-R /note="right homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene" primer_bind complement(8625..8645) /label=T3 /note="T3 promoter, forward primer" promoter complement(8625..8643) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(8664..8680) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 8688..8704 /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(8712..8742) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8757..8778 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 61 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 121 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 181 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 241 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 301 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 361 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 421 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 481 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 541 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 601 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 661 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 721 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 781 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 841 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 901 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 961 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 1021 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 1081 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 1141 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 1201 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 1261 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 1321 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 1381 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 1441 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 1501 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 1561 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 1621 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 1681 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 1741 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 1801 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 1861 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 1921 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa 1981 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc 2041 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga 2101 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag 2161 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt 2221 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta 2281 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag 2341 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg 2401 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg 2461 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg 2521 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg 2581 ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggtacctgc tttctctgac 2641 cagcattctc tcccctgggc ctgtgccgct ttctgtctgc agcttgtggc ctgggtcacc 2701 tctacggctg gcccagatcc ttccctgccg cctccttcag gttccgtctt cctccactcc 2761 ctcttcccct tgctctctgc tgtgttgctg cccaaggatg ctctttccgg agcacttcct 2821 tctcggcgct gcaccacgtg atgtcctctg agcggatcct ccccgtgtct gggtcctctc 2881 cgggcatctc tcctccctca cccaacccca tgccgtcttc actcgctggg ttcccttttc 2941 cttctccttc tggggcctgt gccatctctc gtttcttagg atggccttct ccgacggatg 3001 tctcccttgc gtcccgcctc cccttcttgt aggcctgcat catcaccgtt tttctggaca 3061 accccaaagt accccgtctc cctggcttta gccacctctc catcctcttg ctttctttgc 3121 ctggacaccc cgttctcctg tggattcggg tcacctctca ctcctttcat ttgggcagct 3181 cccctacccc ccttacctct ctagtctgtg ctagctcttc cagccccctg tcatggcatc 3241 ttccaggggt ccgagagctc agctagtctt cttcctccaa cccgggcccc tatgtccact 3301 tcaggacagc atgtttgctg cctccaggga tcctgtgtcc ccgagctggg accaccttat 3361 attcccaggg ccggttaatg tggctctggt tctgggtact tttatctgtc ccctccaccc 3421 cacagtgggg caagcttatt accctgttat ccctaaccgg ttgcttgcaa aagcagtaat 3481 tggaaagcac tctcaaagaa tccaagcttc tgacctcttc tcttcctccc acagggcctc 3541 gagacaattg catcgatggt accagtgaaa gtctgcaatg aattcgagtt ggcgcgcctg 3601 tcattctaaa tctctctttc agcctaaagc tttttccccg tatcccccca ggtgtctgca 3661 ggctcaaaga gcagcgagaa gcgttcagag gaaagcgatc ccgtgccacc ttccccgtgc 3721 ccgggctgtc cccgcacgct gccggctcgg ggatgcgggg ggagcgccgg accggagcgg 3781 agccccgggc ggctcgctgc tgccccctag cgggggaggg acgtaattac atccctgggg 3841 gctttgggag ggggctgtcc ccgtgagctc ccaggcgcgc ctgtcattct aaatctctct 3901 ttcagcctaa agctttttcc ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga 3961 gaagcgttca gaggaaagcg atcccgtgcc accttccccg tgcccgggct gtccccgcac 4021 gctgccggct cggggatgcg gggggagcgc cggaccggag cggagccccg ggcggctcgc 4081 tgctgccccc tagcggggga gggacgtaat tacatccctg ggggctttgg gggggggctg 4141 tccccgtgag ctcccaggcg cgccaactcg gtaccgtatc gatgtcgacg ttaacgctag 4201 tgatatcaac tttgtataga aaagttggct ccgaatttct cgagttaatt aaatgtgctg 4261 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg 4321 ccagtgaatt gtaatacgac tcactatagg gcgaattgga gctaggccta cgtagcgcgc 4381 gagctccacc gcggtggact agttattaat agtaatcaat tacggggtca ttagttcata 4441 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 4501 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 4561 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 4621 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 4681 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 4741 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca 4801 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag 4861 cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg 4921 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt 4981 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg 5041 ggagtcgagt cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc 5101 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc 5161 ctccgggctg taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg 5221 aaagccttaa agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg 5281 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc 5341 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg 5401 gggcggtgcc ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg 5461 tgcgtggggg ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca 5521 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtgcggggcg 5581 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc 5641 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccg gagcgccggc 5701 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg 5761 cagggacttc ctttgtccca aatctggcgg agccgaaatc tgggaggcgc cgccgcaccc 5821 cctctagcgg gcgcgggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg 5881 ccttcgtgcg tcgccgcgcc gccgtcccct tctccatctc cagcctcggg gctgccgcag 5941 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc 6001 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg 6061 ggcaacgtgc tggttgttgt gctgtctcat cattttggca aagaattcga cccaagtttg 6121 tacaaaaaag caggctgaat ccaccggtcg ccaccatggt gagcaagggc gaggagctgt 6181 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca 6241 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct 6301 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc 6361 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca 6421 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga 6481 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca 6541 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc 6601 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc 6661 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca 6721 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga 6781 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg 6841 ggatcactct cggcatggac gagctgtaca agtaataccc agctttcttg tacaaagtgg 6901 tacgcgtgaa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc 6961 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga 7021 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc 7081 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca 7141 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc 7201 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt 7261 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt 7321 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt 7381 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct 7441 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 7501 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 7561 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 7621 gaaacctgtc gtgccagcgg atcgacagta ctaagcttgg tagataagtt gatttagcca 7681 taaaatattg tttccgtgac cccttaatta atagggataa cagggtaatc tcgaggttta 7741 aacgtcgacg cggccgctac tagggacagg attggtgaca gaaaagcccc atccttaggc 7801 ctcctccttc ctagtctcct gatattgggt ctaaccccca cctcctgtta ggcagattcc 7861 ttatctggtg acacaccccc atttcctgga gccatctctc tccttgccag aacctctaag 7921 gtttgcttac gatggagcca gagaggatcc tgggagggag agcttggcag ggggtgggag 7981 ggaagggggg gatgcgtgac ctgcccggtt ctcagtggcc accctgcgct accctctccc 8041 agaacctgag ctgctctgac gcggctgtct ggtgcgtttc actgatcctg gtgctgcagc 8101 ttccttacac ttcccaagag gagaagcagt ttggaaaaac aaaatcagaa taagttggtc 8161 ctgagttcta actttggctc ttcacctttc tagtccccaa tttatattgt tcctccgtgc 8221 gtcagtttta cctgtgagat aaggccagta gccagccccg tcctggcagg gctgtggtga 8281 ggaggggggt gtccgtgtgg aaaactccct ttgtgagaat ggtgcgtcct aggtgttcac 8341 caggtcgtgg ccgcctctac tccctttctc tttctccatc cttctttcct taaagagtcc 8401 ccagtgctat ctgggacata ttcctccgcc cagagcaggg tcccgcttcc ctaaggccct 8461 gctctgggct tctgggtttg agtccttggc aagcccagga gaggcgctca ggcttccctg 8521 tcccccttcc tcgtccacca tctcatgccc ctggctctcc tgccccttcc ctacaggggt 8581 tcctggctct gctctccacc gcggtggagc tccagctttt gttcccttta gtgagggtta 8641 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 8701 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 8761 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 8821 tcgtgccagc tgcattaatg aatc //