LOCUS Exported 5164 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV vector expressing Cre from rTH promoter. ACCESSION . VERSION . KEYWORDS AAV.rTH.PI.Cre.SV40 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5164) TITLE AAV plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 5164) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5164 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 63..84 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 99..129 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 137..153 /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)." primer_bind 142..164 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 161..177 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 161..177 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" repeat_region 215..344 /label=AAV2 ITR intron 817..949 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 932..951 /label=pMT2-F /note="Synthetic intron, forward primer" promoter 994..1012 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1061..2092 /codon_start=1 /product="site-specific recombinase" /label=Cre /note="Cre is a site-specific recombinase from bacteriophage P1. Recombination occurs at loxP sequences." /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE DGD" polyA_signal 2145..2266 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2156..2175) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2210..2229 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 2428..2557 /label=AAV2 ITR primer_bind complement(2576..2593) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2576..2592) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2585..2607) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2734..3189 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2821..2840) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3031..3052 /label=F1ori-F /note="F1 origin, forward primer" promoter 3215..3319 /gene="bla" /label=AmpR promoter CDS 3320..4180 /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 complement(3538..3557) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4351..4939 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4840..4859 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5093..5110 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca 61 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct 121 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat 181 gattacgcca gatttaatta aggccttaat taggctgcgc gctcgctcgc tcactgaggc 241 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg 301 agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa 361 cccgccatgc tacttatcta cgtagccatg ctctaggaag atcaattcct agcggtctcc 421 tgtcccacag aataccagcc agcccctgcc ctacgtcgtg cctcgggctg agggtgattc 481 agaggcaggt gcctgtgaca gtggatgcaa ttagatctaa tgggacggag gcctttctcg 541 tcgccctcgc tccatgccca cccccgcctc cctcaggcac agcaggcgtg gagaggatgc 601 gcaggaggta ggaggtgggg gacccagagg ggctttgacg tcagcctggc ctttaaagag 661 ggcgcctgcc tggcgagggc tgtggagaca gaactcggga ccaccagaag ctttattgcg 721 gtagtttatc acagttaaat tgctaacgca gtcagtgctt ctgacacaac agtctcgaac 781 ttaagctgca gaagttggtc gtgaggcact gggcaggtaa gtatcaaggt tacaagacag 841 gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt gcgtttctga 901 taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg tgtccactcc 961 cagttcaatt acagctctta aggctagagt acttaatacg actcactata ggctagtcga 1021 gaattgtaca ctttacttaa aaccattatc tgagtgtgaa atgtccaatt tactgaccgt 1081 acaccaaaat ttgcctgcat taccggtcga tgcaacgagt gatgaggttc gcaagaacct 1141 gatggacatg ttcagggatc gccaggcgtt ttctgagcat acctggaaaa tgcttctgtc 1201 cgtttgccgg tcgtgggcgg catggtgcaa gttgaataac cggaaatggt ttcccgcaga 1261 acctgaagat gttcgcgatt atcttctata tcttcaggcg cgcggtctgg cagtaaaaac 1321 tatccagcaa catttgggcc agctaaacat gcttcatcgt cggtccgggc tgccacgacc 1381 aagtgacagc aatgctgttt cactggttat gcggcggatc cgaaaagaaa acgttgatgc 1441 cggtgaacgt gcaaaacagg ctctagcgtt cgaacgcact gatttcgacc aggttcgttc 1501 actcatggaa aatagcgatc gctgccagga tatacgtaat ctggcatttc tggggattgc 1561 ttataacacc ctgttacgta tagccgaaat tgccaggatc agggttaaag atatctcacg 1621 tactgacggt gggagaatgt taatccatat tggcagaacg aaaacgctgg ttagcaccgc 1681 aggtgtagag aaggcactta gcctgggggt aactaaactg gtcgagcgat ggatttccgt 1741 ctctggtgta gctgatgatc cgaataacta cctgttttgc cgggtcagaa aaaatggtgt 1801 tgccgcgcca tctgccacca gccagctatc aactcgcgcc ctggaaggga tttttgaagc 1861 aactcatcga ttgatttacg gcgctaagga tgactctggt cagagatacc tggcctggtc 1921 tggacacagt gcccgtgtcg gagccgcgcg agatatggcc cgcgctggag tttcaatacc 1981 ggagatcatg caagctggtg gctggaccaa tgtaaatatt gtcatgaact atatccgtaa 2041 cctggatagt gaaacagggg caatggtgcg cctgctggaa gatggcgatt agccattaac 2101 gcggcgtggt acctctagag tcgggccgct tcgagcagac atgataagat acattgatga 2161 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga 2221 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg 2281 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa 2341 cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt 2401 agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct 2461 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct 2521 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagcct taattaacct aattcactgg 2581 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg 2641 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt 2701 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg 2761 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 2821 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 2881 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 2941 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc 3001 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac 3061 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt 3121 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc 3181 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 3241 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 3301 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt 3361 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc 3421 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat 3481 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct 3541 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca 3601 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg 3661 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa 3721 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg 3781 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga 3841 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg 3901 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt 3961 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg 4021 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc 4081 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca 4141 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc 4201 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 4261 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 4321 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 4381 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 4441 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct 4501 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 4561 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 4621 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 4681 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 4741 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 4801 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 4861 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 4921 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 4981 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 5041 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 5101 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc 5161 gatt //