LOCUS       sequence-389701-        6160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The AAV vector for iRFP713(V256C) and EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6160)
  TITLE     Plasmids for KOBACHI mice project
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6160)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-CMV-iRFP713(V256C)-P2A-EGFP
FEATURES             Location/Qualifiers
     source          1..6160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(206..310)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(336..791)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(473..494)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     685..704
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   863..992
                     /label=AAV2 ITR
     enhancer        1125..1428
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1429..1632
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1583..1603
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1629..1653
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1735..1744
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2701..2757
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(2767..2769,2770..2772,2773..3285)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(2812..2833)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3073..3092)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3420..3441
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3466..4054
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3519..3539)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3937..3948)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4097..4218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4108..4127)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4162..4181
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4380..4509
                     /label=AAV2 ITR
     rep_origin      complement(4746..5334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4826..4845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5505..6160,1..136,137..205))
                     /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     6128..6147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
       61 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
      121 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
      181 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
      241 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
      301 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
      361 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
      421 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
      481 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
      541 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
      601 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
      661 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
      721 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
      781 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
      841 ggcctcttcg ctattacgcc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
      901 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
      961 gggagtggcc aactccatca ctaggggttc cttgtagtta atgattaacc cgccatgcta
     1021 cttatctacg tagccatgct ctaggaagat cgtaggttaa ttaatagtta ttaatagtaa
     1081 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     1141 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     1201 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     1261 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     1321 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     1381 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     1441 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     1501 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     1561 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     1621 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     1681 gacctccata gaagacaccg ggaccgatcc agcctccgcg gattcgaaac tagtgccacc
     1741 atggccgaag ggagcgtggc aagacagccc gatctgctga catgcgacga cgaaccaatc
     1801 cacattccag gagctatcca gcctcatgga ctgctgctgg ctctggccgc tgacatgact
     1861 atcgtggcag gctctgataa cctgccagag ctgaccgggc tggctatcgg agcactgatt
     1921 ggccggtccg cagccgacgt gttcgattct gagactcaca atagactgac catcgccctg
     1981 gctgaacctg gggctgcagt gggagcccca attaccgtcg gcttcacaat gcgaaaggac
     2041 gctggcttta tcgggtcctg gcaccgccat gatcagctga tttttctgga gctggaacca
     2101 cctcagagag acgtggcaga gccacaggct ttctttcgga gaacaaacag cgccatcagg
     2161 cgcctgcagg ccgctgagac tctggaatcc gcttgcgcag ccgctgcaca ggaagtgagg
     2221 aagatcaccg ggttcgatcg cgtcatgatc tacagattcg cctcagactt tagcggagaa
     2281 gtgatcgcag aagatcgctg tgccgaggtc gaaagtaaac tgggcctgca ttacccagcc
     2341 tcaaccgtgc ctgcacaggc acgacggctg tatacaatca accccgtgcg gatcattccc
     2401 gacattaatt acagaccagt gcccgtcaca cctgacctga acccagtgac tgggaggccc
     2461 atcgatctga gcttcgccat tctgcgcagc gtgtcccctt gtcacctgga attcatgagg
     2521 aatatcggaa tgcatggcac catgtctatc agtattctga gaggcgaaag gctgtggggg
     2581 ctgatcgtgt gccaccatcg gacaccttac tatgtggacc tggacggcag gcaggcttgt
     2641 gaactggtgg ctcaggtgct ggcttggcag attggcgtga tggaactcga gggaagcgga
     2701 gcgacgaatt ttagtctact gaaacaagcg ggagacgtgg aggaaaaccc tggacctggc
     2761 ggccgcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2821 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2881 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2941 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     3001 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     3061 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     3121 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3181 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     3241 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     3301 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3361 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     3421 atggtcctgc tggagttcgt gaccgccgcc tagaagctta tcgataatca acctctggat
     3481 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3541 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3601 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3661 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3721 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3781 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3841 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     3901 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3961 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     4021 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg
     4081 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4141 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4201 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     4261 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     4321 gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     4381 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4441 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4501 agcgcgcagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4561 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4621 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4681 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4741 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4801 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4861 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4921 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4981 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5041 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5101 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5161 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5221 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5281 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5341 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5401 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5461 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5521 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5581 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5641 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5701 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5761 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5821 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5881 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5941 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6001 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6061 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6121 atacgggata ataccgcgcc acatagcaga actttaaaag
//
