LOCUS Exported 6304 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Flp-dependent mCherry Control. ACCESSION . VERSION . KEYWORDS pAAV-Ef1a-fDIO mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6304) TITLE mCherry Control JOURNAL Unpublished REFERENCE 2 (bases 1 to 6304) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6304 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 231..1409 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 462..1400 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1357..1377 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 1440..1473 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" CDS complement(1566..2276) /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(1844..1863) /label=mCherry-F /note="mCherry, forward primer" primer_bind 2113..2131 /label=mCherry-R /note="mCherry, reverse primer" regulatory 2273..2282 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(2289..2322) /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" misc_feature 2431..3019 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2484..2504) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2902..2913) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3051..3527 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3250..3269) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3567..3707 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3567..3696 /label=AAV2 ITR rep_origin 3782..4237 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3869..3888) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4079..4100 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4254..4273) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4373..4395 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4433..4451) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4519..4623 /gene="bla" /label=AmpR promoter CDS 4624..5484 /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(4842..4861) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5655..6243 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6144..6163 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga 181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg 241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg 301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt 361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg 421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt 481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct 541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga 601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct 661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt 721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc 781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg 841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac 901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc 961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag 1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct 1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg 1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga 1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc 1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg 1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa 1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggag 1441 aagttcctat tctctagaaa gtataggaac ttcgcagaat ggtagctgga ttgtagctgc 1501 tattagcaat atgaaacctc ttagaagttc ctattcttca aaaggtatag gaacttcggc 1561 gcgccttact tgtacagctc gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg 1621 tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt gatgttgacg 1681 ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcagcg 1741 tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt cagggcgccg 1801 tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt ctgcattacg 1861 gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc 1921 tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc 1981 tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc 2041 ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc 2101 agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc 2161 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg 2221 aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct caccatggtg 2281 gcgctagcga agttcctata ctttctagag aataggaact tcttgcctta acccagaaat 2341 tatcactgtt attctttaga atggtgcaaa gagaagttcc tatacctttt gaagaatagg 2401 aacttcgaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa 2461 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta 2521 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa 2581 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg 2641 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc 2701 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc 2761 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg 2821 gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg 2881 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg 2941 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc 3001 ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc 3061 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca 3121 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata 3181 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg 3241 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa 3301 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc 3361 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca 3421 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca 3481 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg 3541 taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc 3601 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt 3661 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg 3721 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag 3781 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 3841 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 3901 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 3961 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg 4021 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 4081 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta 4141 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt 4201 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc 4261 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc 4321 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc 4381 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg 4441 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc 4501 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat 4561 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag 4621 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt 4681 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt 4741 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc 4801 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta 4861 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac 4921 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa 4981 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg 5041 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc 5101 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg 5161 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta 5221 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg 5281 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg 5341 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc 5401 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt 5461 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt 5521 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc 5581 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag 5641 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 5701 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 5761 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag 5821 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 5881 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 5941 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 6001 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 6061 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 6121 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 6181 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 6241 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 6301 atgt //