LOCUS Exported 7454 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS rAAV-syn::FLEX-rev::PSAML141F,Y115F:GlyR-IRES-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7454) AUTHORS Magnus CJ, Lee PH, Atasoy D, Su HH, Looger LL, Sternson SM TITLE Chemical and genetic engineering of selective ion channel-ligand interactions. JOURNAL Science. 2011 Sep 2;333(6047):1292-6. PUBMED 21885782 REFERENCE 2 (bases 1 to 7454) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7454 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 55..184 /label=AAV2 ITR intron 777..909 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 892..911 /label=pMT2-F /note="Synthetic intron, forward primer" protein_bind complement(922..955) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(1018..1051) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(1062..1778) /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(1107..1128) /label=EGFP-C /note="EGFP, forward primer" primer_bind 1456..1475 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1715..1736 /label=EGFP-N /note="EGFP, reverse primer" misc_feature 1802..2353 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind 2192..2209 /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" protein_bind 3708..3741 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 3812..3845 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3917..4505 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3970..3990) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4388..4399) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4583..4704 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4594..4613) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4648..4667 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 4866..4995 /label=AAV2 ITR primer_bind complement(5061..5078) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5232..5820) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5312..5331) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5991..6851) /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 6614..6633 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6852..6956) /gene="bla" /label=AmpR promoter rep_origin complement(6982..7437) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7119..7140) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 7331..7350 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctgcgc 61 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc 121 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt 181 tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag 241 atccgagctc cagtgtgctg gaattcgccc ttttcagtat ttaaattgac gaccgacccc 301 gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg 361 gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca 421 gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg 481 acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc 541 gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac 601 catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg cagcggagga 661 gtcgtgtcgt gcctgagagc gcagctgtgc tcctgggcac cgcgcagtcc gcccccgcgg 721 ctcctggcca gaccacccct aggaccccct gccccaagtc gcagccgttg gatcaggtaa 781 gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga 841 gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc 901 tctccacagg tgatgatccc cataacttcg tataaagtat cctatacgaa gttatatcaa 961 aataggaaga ccaatgcttc accatcgacc cgaattgcca agcatcacca tcgacacata 1021 acttcgtata atgtatgcta tacgaagtta tactagctag attacttgta cagctcgtcc 1081 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc 1141 tcgttggggt ctttgctcag cttggactgg gtgctcaggt agtggttgtc gggcagcagc 1201 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg 1261 tcctcgatgt tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc 1321 atgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg 1381 tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac 1441 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg 1501 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag 1561 cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc 1621 agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg 1681 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg 1741 ggcaccaccc cggtgaacag ctcctcgccc ttgctcaccg gatcccccat ggttgtggcc 1801 atattatcat cgtgtttttc aaaggaaaac cacgtccccg tggttcgggg ggcctagacg 1861 tttttttaac ctcgactaaa cacatgtaaa gcatgtgcac cgaggcccca gatcagatcc 1921 catacaatgg ggtaccttct gggcatcctt cagccccttg ttgaatacgc ttaggagagc 1981 catttgactc tttccacaac tatccaactc acaacgtggc actggggttg tgccgccttt 2041 gcaggtgtat cttatacacg tggcttttgg ccgcagaggc acctgtcgcc aggtgggggg 2101 ttccgctgcc tgcaaagggt cgctacagac gttgtttgtc ttcaagaagc ttccagagga 2161 actgcttcct tcacgacatt caacagacct tgcattcctt tggcgagagg ggaaagaccc 2221 ctaggaatgc tcgtcaagaa gacagggcca ggtttccggg ccctcacatt gccaaaagac 2281 ggcaatatgg tggaaaataa catatagaca aacgcacacc ggccttattc caagcggctt 2341 cggccagtaa cgttaggggg gggggagggg ggaattgcgg ccgcttactg gttgtgtacg 2401 tcctcccttc gcacaatctt gtagatgatc caataaaaca tgttgaaaat caggaaagcc 2461 atggggaatc cgatccggga gattttgtcg atctttttcg ctctctgaat aaagagtttg 2521 cgcatttcct ccgggctctt agatggagca gggggtgggt tggttgtgtt gctattgttg 2581 gctcccttca cggaaattcc gtctttagcc tggaggcaag caggtcccat accataggcg 2641 ctaaagttga agcgtccttc tcctgcctca tcctctttgt ggtggcgtcg ttttcttctg 2701 aaccgcaaca gctctttatg ctgtcgagac acaaaattga cagcggcgta ttccagcagt 2761 gcgctgaaga caaacagaag gcagacagcc atccagatat cgatggcctt cacgtaagaa 2821 accttgggca agctggctcg gctgcctgag ctctgagtag tcattgtcag cactgtggtg 2881 atgcccagtc cgaccctagc tggagcggcg tccatattaa tccagaagga gatccatgac 2941 aaaatcacta taagcaagct tgggatatac atttggatca gataataacc catgcgtctc 3001 cgcatggtca ctgtgaaggt gacatctggg tagggctctt tgcagcattc ataaaatcgc 3061 tcgcttcttt taccggggat tcccacaagg tcccactccc cattcgggat gtaaccagag 3121 atgtcagcct cctgcatttg cagatccagg ctccatccgc cgtagctcca gctcccgaat 3181 ttgagtttgc agtgctggac gtcgaaggga aaccagcgga cgtcgatata gcatgaggac 3241 ttgaagattc caggggggaa gtactggcag tggcctgaag aattgaccaa cacgttggtg 3301 tgaaatgttg catcaaacct ttcgtctgcg ctgttgaaca gcaggatatc tggtttccag 3361 atttggccgt cagggaatct tacagtcttt acacctgggt attcgctcac attccactgc 3421 aagtagtgat cggtccagga catctgcaac caaatattag tggtcagcac ctggttcttt 3481 tcatccacgt ccatgatctg cagcaaggag aggctgaagt acactgtcag tggctgggaa 3541 tcatttgcga ccggccgctc cagggggtta taattcttaa caagctcctt atacagtttg 3601 cgctggaact caccctgcag gctaacgtgc agaagggaag cagccagggc gagccacacg 3661 ccgcctggag aacagcgcat ggtgaattca agcttgctag ccctagtata acttcgtata 3721 ggatacttta tacgaagtta tcattgggat tcttcctatt ttgatccaag catcaccatc 3781 gaccctctag tccagatctc accatcgacc cataacttcg tatagcatac attatacgaa 3841 gttatgtccc tcgaagaggt tcgaaacact gaaaagggcg aattctgcag atgatccact 3901 cgagaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat 3961 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca 4021 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc 4081 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc 4141 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt 4201 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg 4261 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc 4321 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta 4381 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg 4441 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc 4501 cccgcatcga taccgtcgac ccgggcggcc gctcgagcat gcatctagag tcgacccggg 4561 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga 4621 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc 4681 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt 4741 cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc 4801 gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa 4861 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac 4921 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag 4981 cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 5041 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 5101 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 5161 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 5221 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 5281 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 5341 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 5401 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 5461 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 5521 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 5581 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 5641 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 5701 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 5761 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 5821 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 5881 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 5941 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 6001 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 6061 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 6121 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 6181 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 6241 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 6301 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 6361 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 6421 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 6481 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 6541 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 6601 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 6661 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 6721 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 6781 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 6841 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 6901 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 6961 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa 7021 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata 7081 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac 7141 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc 7201 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa 7261 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg 7321 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg 7381 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc 7441 attcgccatt cagg //