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synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5164) TITLE AAV plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 5164) AUTHORS .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" repeat_region 1..130 /label=AAV2 ITR intron 603..735 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 718..737 /label=pMT2-F /note="Synthetic intron, forward primer" promoter 780..798 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 847..1878 /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 1931..2052 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1942..1961) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1996..2015 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 2214..2343 /label=AAV2 ITR primer_bind complement(2362..2379) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2362..2378) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2371..2393) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2520..2975 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2607..2626) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2817..2838 /label=F1ori-F /note="F1 origin, forward primer" promoter 3001..3105 /gene="bla" /label=AmpR promoter CDS 3106..3966 /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(3324..3343) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4137..4725 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4626..4645 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4879..4896 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5013..5034 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5049..5079 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5087..5103 /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 5092..5114 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5111..5127 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5111..5127 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct 181 aggaagatca attcctagcg gtctcctgtc ccacagaata ccagccagcc cctgccctac 241 gtcgtgcctc gggctgaggg tgattcagag gcaggtgcct gtgacagtgg atgcaattag 301 atctaatggg acggaggcct ttctcgtcgc cctcgctcca tgcccacccc cgcctccctc 361 aggcacagca ggcgtggaga ggatgcgcag gaggtaggag gtgggggacc cagaggggct 421 ttgacgtcag cctggccttt aaagagggcg cctgcctggc gagggctgtg gagacagaac 481 tcgggaccac cagaagcttt attgcggtag tttatcacag ttaaattgct aacgcagtca 541 gtgcttctga cacaacagtc tcgaacttaa gctgcagaag ttggtcgtga ggcactgggc 601 aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga 661 gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg 721 cctttctctc cacaggtgtc cactcccagt tcaattacag ctcttaaggc tagagtactt 781 aatacgactc actataggct agtcgagaat tgtacacttt acttaaaacc attatctgag 841 tgtgaaatgt ccaatttact gaccgtacac caaaatttgc ctgcattacc ggtcgatgca 901 acgagtgatg aggttcgcaa gaacctgatg gacatgttca gggatcgcca ggcgttttct 961 gagcatacct ggaaaatgct tctgtccgtt tgccggtcgt gggcggcatg gtgcaagttg 1021 aataaccgga aatggtttcc cgcagaacct gaagatgttc gcgattatct tctatatctt 1081 caggcgcgcg gtctggcagt aaaaactatc cagcaacatt tgggccagct aaacatgctt 1141 catcgtcggt ccgggctgcc acgaccaagt gacagcaatg ctgtttcact ggttatgcgg 1201 cggatccgaa aagaaaacgt tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa 1261 cgcactgatt tcgaccaggt tcgttcactc atggaaaata gcgatcgctg ccaggatata 1321 cgtaatctgg catttctggg gattgcttat aacaccctgt tacgtatagc cgaaattgcc 1381 aggatcaggg ttaaagatat ctcacgtact gacggtggga gaatgttaat ccatattggc 1441 agaacgaaaa cgctggttag caccgcaggt gtagagaagg cacttagcct gggggtaact 1501 aaactggtcg agcgatggat ttccgtctct ggtgtagctg atgatccgaa taactacctg 1561 ttttgccggg tcagaaaaaa tggtgttgcc gcgccatctg ccaccagcca gctatcaact 1621 cgcgccctgg aagggatttt tgaagcaact catcgattga tttacggcgc taaggatgac 1681 tctggtcaga gatacctggc ctggtctgga cacagtgccc gtgtcggagc cgcgcgagat 1741 atggcccgcg ctggagtttc aataccggag atcatgcaag ctggtggctg gaccaatgta 1801 aatattgtca tgaactatat ccgtaacctg gatagtgaaa caggggcaat ggtgcgcctg 1861 ctggaagatg gcgattagcc attaacgcgg cgtggtacct ctagagtcgg gccgcttcga 1921 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa 1981 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc 2041 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg 2101 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt 2161 cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc 2221 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg 2281 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg 2341 cagccttaat taacctaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc 2401 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag 2461 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga 2521 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 2581 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 2641 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 2701 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 2761 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 2821 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 2881 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 2941 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg 3001 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 3061 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat 3121 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca 3181 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc 3241 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca 3301 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg 3361 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca 3421 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata 3481 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag 3541 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg 3601 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca 3661 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta 3721 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct 3781 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca 3841 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag 3901 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat 3961 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt 4021 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 4081 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 4141 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 4201 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 4261 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 4321 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 4381 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 4441 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 4501 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 4561 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 4621 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 4681 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 4741 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 4801 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 4861 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc 4921 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc 4981 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca 5041 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa 5101 caatttcaca caggaaacag ctatgaccat gattacgcca gatttaatta aggccttaat 5161 tagg //