LOCUS Exported 7037 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS AAV-EF1a-BbTagBY SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7037) AUTHORS Cai D, Cohen KB, Luo T, Lichtman JW, Sanes JR TITLE Improved tools for the Brainbow toolbox. JOURNAL Nat Methods. 2013 May 5;10(6):540-7. doi: 10.1038/nmeth.2450. Epub 2013 May 5. PUBMED 23817127 REFERENCE 2 (bases 1 to 7037) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7037 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 12..141 /label=AAV2 ITR promoter 242..1419 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 473..1410 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1367..1387 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 1426..1459 /label=lox71 /bound_moiety="Cre recombinase" /note="Left element (LE) mutant of loxP (Araki et al., 2010). Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 2230..2239 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(2240..2273) /label=loxKR3 /bound_moiety="Cre recombinase" /note="Right element (RE) mutant of loxP (Araki et al., 2010). Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 2277..2310 /label=lox66 /bound_moiety="Cre recombinase" /note="Right element (RE) mutant of loxP (Araki et al., 2010). Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(2389..3105) /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2431..2452) /label=EGFP-C /note="EGFP, forward primer" primer_bind 2780..2799 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3039..3060 /label=EGFP-N /note="EGFP, reverse primer" regulatory 3102..3111 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(3112..3145) /label=loxKR3 /bound_moiety="Cre recombinase" /note="Right element (RE) mutant of loxP (Araki et al., 2010). Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3164..3752 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3217..3237) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3635..3646) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3784..4260 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3983..4002) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4300..4440 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4300..4429 /label=AAV2 ITR rep_origin 4515..4970 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4602..4621) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4812..4833 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4987..5006) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5106..5128 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5166..5184) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5252..5356 /gene="bla" /label=AmpR promoter CDS 5357..6217 /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(5575..5594) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6388..6976 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6877..6896 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt 181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat 241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg 301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag 361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc 421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc 481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat 541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtacctaccg ttcgtatagc 1441 atacattata cgaagttatt taggagagca cacacttgca gctcatgcag ccggggccac 1501 tctcatcagg agggttcagc ttactcacgc gtgcacttct gtgccccagt ttgctaggga 1561 ggtcgcagta tctggccact gccacctcgt gctgctcgac gtaggtctcg ttgttggcct 1621 ccttgattct ttccagtctg tagtccacat agtagacgcc aggcatcttg aggttcttag 1681 cgggtttctt ggatctatat gtggtcttga tgtttgcgat cagatggctc ccgcccacga 1741 gcttcagggc catgtcgttt ctgccttcca ggccgccgtc agcggggtac agcgtctcgg 1801 tgaaggcctc ccagccgagt gttttcttct gcatcacagg gccgttggat gtgaagttca 1861 cccctctgat cttgacgttg tagatgaggc agccgtcctg gaggctggtg tcctgggtag 1921 cggtcagcac gcccccgtct tcgtatgtgg tgactctctc ccatgtgaag ccctcaggga 1981 aggactgctt gaagaagtcg gggatgccct gggtgtggtt gatgaaggtc ttgctgccgt 2041 agaggaagct agtagccagg atgtcgaagg cgaaggggag agggccgccc tcgaccacct 2101 tgattctcat ggtctgggtg ccctcgtagg gcttgccttc gccctcggat gtgcacttga 2161 agtgatggtt gtccacggtg ccctccatgt acagcttcat gtgcatgttc tccttaatca 2221 gctcgctcac catggtggca taacaaggta taatgtatgc tatacgaagt tatgatataa 2281 cttcgtatag catacattat acgaacggta tcaggagagc acacacttgc agctcatgca 2341 gccggggcca ctctcatcag gagggttcag cttactcacg cgtgcactct tgtacagctc 2401 gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg 2461 cttctcgttg gggtctttgc tcagggcgga ctggtagctc aggtagtggt tgtcgggcag 2521 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct 2581 gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc 2641 ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt 2701 gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc 2761 gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc 2821 ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg 2881 gtagcgggcg aagcactgca ggccgtagcc gaaggtggtc acgagggtgg gccagggcac 2941 gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc 3001 ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag 3061 gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg cataacaagg 3121 tataatgtat gctatacgaa gttatgatat caagcttatc gataatcaac ctctggatta 3181 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg 3241 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc 3301 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca 3361 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac 3421 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact 3481 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc 3541 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg 3601 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc 3661 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac 3721 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc 3781 tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact 3841 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg 3901 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag 3961 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct 4021 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag 4081 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga 4141 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca 4201 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt 4261 ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt 4321 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc 4381 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg 4441 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc 4501 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 4561 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 4621 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 4681 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg 4741 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 4801 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta 4861 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 4921 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac 4981 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc 5041 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac 5101 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg 5161 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta 5221 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 5281 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 5341 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 5401 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 5461 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 5521 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 5581 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 5641 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 5701 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 5761 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 5821 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 5881 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga 5941 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 6001 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 6061 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 6121 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 6181 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 6241 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 6301 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 6361 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 6421 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 6481 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct 6541 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 6601 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 6661 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 6721 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 6781 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 6841 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 6901 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 6961 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 7021 gccttttgct cacatgt //