LOCUS       sequence-399512-        6637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Truncated version of human Tyrosinase as control virus for 
            Tyrosinase expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6637)
  TITLE     Prigge Lab AAV vectors
REFERENCE   2  (bases 1 to 6637)
  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"
COMMENT     Sequence Label: pAAV_Ef1a_DIO_hTyrNull_HATag
FEATURES             Location/Qualifiers
     source          1..6637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    25..73
                     /note="synthetic polyadenylation signal"
     repeat_region   103..243
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   103..232
                     /label=AAV2 ITR
     rep_origin      318..773
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(405..424)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     615..636
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(790..809)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     909..931
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(969..987)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1055..1159
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1160..1228,1229..2020)
                     /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(1378..1397)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2191..2779
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2680..2699
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2841..2981
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2852..2981
                     /label=AAV2 ITR
     promoter        3082..4259
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3313..4250
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4207..4227
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(4290..4323)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(4374..4407)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4419..4445)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(6075..6101)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    6122..6155
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    6206..6239
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    join(6252..6637,1..18)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
ORIGIN
        1 gtcggggaag ctgacgtcag cgctaataaa atcgctatcc atcgaagatg gatgtgtgtt
       61 ggttttttgt gtgggtaacc cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
      121 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
      181 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
      241 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
      301 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
      361 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
      421 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
      481 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
      541 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
      601 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg
      661 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
      721 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
      781 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
      841 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
      901 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
      961 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     1021 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1081 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1141 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1201 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1261 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1321 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1381 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     1441 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1501 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1561 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1621 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1681 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     1741 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     1801 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     1861 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     1921 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1981 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     2041 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2101 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2161 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2221 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2281 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2341 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2401 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2461 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2521 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2581 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2641 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2701 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2761 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2821 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     2881 ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     2941 gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca cgcgtaagct
     3001 ttgcaaagat ggataaagtt ttaaacagag aggaatcttt gcagctaatg gaccttctag
     3061 gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc
     3121 acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg
     3181 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     3241 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     3301 gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat
     3361 ggcccttgcg tgccttgaat tacttccact ggctgcagta cgtgattctt gatcccgagc
     3421 ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc
     3481 gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc
     3541 ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg
     3601 ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg
     3661 gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt
     3721 cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg
     3781 gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa
     3841 ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg
     3901 cagggagctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac
     3961 aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg
     4021 cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg
     4081 gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc
     4141 cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt
     4201 tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag
     4261 gtaccggatc ctctagagtc gactccggaa taacttcgta taggatactt tatacgaagt
     4321 tatgcagaat ggtagctgga ttgtagctgc tattagcaat atgaaacctc ttaataactt
     4381 cgtatagcat acattatacg aagttatggc gcgccttaag cgtaatctgg aacatcgtat
     4441 gggtaaaact ccccggaccc agctgctaaa tggctctgat acaagctgtg gtaatcctct
     4501 ttctccatga ggagtggctg cttttcttca ggaagctgct ttctcttgtg acgacacagc
     4561 aagctcacaa gccctgccag cagggcagtg aggacggccc ctaccatcgc cgccccaagg
     4621 agccatgacc agatccgact cgcttgttcc aaataggact taatgtagtc ttgaaaagag
     4681 tctgggtctg aatcttgtag atagctatag tcatagccca gatctttgga tgaaataaag
     4741 aaatcaccat ttctgtacag tggtataaaa ggaaccatgt aggattcccg gttatgtcca
     4801 atgggtgcat tggcttctgg ataaacttct tgaagaggac ggtgccttcg gagccactgc
     4861 tcaaaaatac tgtcaacaaa tgcatggtga agaaggaaga taggatcgtt ggcagatccc
     4921 tgtacctggg acattgttcc attcatatag atgtgcaagg cattgtgcat gctgctttga
     4981 gaggcatccg ctatcccagt aagtggacta gcaaatcctt ccagtgtatt tctaaagctg
     5041 aaattggcag ctttatccat ggaaccagat tcatattggg tcaaactcag gcaaaattct
     5101 acatcagctg aagaggggag ccttggggtt ctggatttgt catggtttcc aggattacgc
     5161 cgtaaaggtc cctcgggcgt tccattgcat aaagactgat ggctgttgta ctcctccaat
     5221 cggctacaga caatctgcca agaggagaag aatgatgctg ggctgagtaa gttaggattt
     5281 gtggggtgct gacctcccat gtactcatct gtgcaaatgt cacacttttc tgcatcccgc
     5341 cagtcccaat atggaatagt gaagttttca tctcctgtca gcttctggat ttcttgttcc
     5401 caccgcaaca agaagagtct atgccaaggc agaaaagctg gtgcttcatg ggcaaaatca
     5461 atgtctctcc agatttcaga tcccccaagc agtgcatcca ttgacacata ataatgttac
     5521 atccagacaa agaggtcata aatattgatg tcgttaaaca tgggtgttga tccatttttc
     5581 atttggccat aggtccctat ggggatgaca tagtctgagc tgatggtatg ctttgctaaa
     5641 gtgaggtagg caaaaaattt gtccttctct ggggcactca aatcgaagat gtttcttctc
     5701 accaagagtc gtctctctgt gcagtttggt ccccaaaagc caaacttgca gtttccacag
     5761 ttgaatccca tgaagttgcc agagcactgg caggtcctat tataaaagac ggaaggccac
     5821 gactcccggt catccacccc tgtgaaggga aattgaggcc caagtggtgc attggacaga
     5881 aggatattct gacaggaacc tctgcctgaa agctggccac agggactcct gtccccgctc
     5941 cacggtggac agcattcctt ctccatcagg ttcttagagg agacacaggc tctagggaaa
     6001 tggccagcgg aggtctggaa actccacagc aggcagtaca aaacagccag gagaaactcc
     6061 ccggacccag ctgcagcgta atctggaaca tcgtatgggt acatcaaggt cttaggctag
     6121 cataacttcg tataaagtat cctatacgaa gttatttgcc ttaacccaga aattatcact
     6181 gttattcttt agaatggtgc aaagaataac ttcgtataat gtatgctata cgaagttatg
     6241 aattcaagct taatcaacct ctggattaca aaatttgtga aagattgact gatattctta
     6301 actatgttgc tccttttacg ctgtgtggat atgctgcttt aatgcctctg tatcatgcta
     6361 ttgcttcccg tatggctttc gttttctcct ccttgtataa atcctggttg ctgtctcttt
     6421 atgaggagtt gtggcccgtt gtccgtcaac gtggcgtggt gtgctctgtg tttgctgacg
     6481 caacccccac tggctggggc attgccacca cctgtcaact cctttctggg actttcgctt
     6541 tccccctccc gatcgccacg gcagaactca tcgccgcctg ccttgcccgc tgctggacag
     6601 gggctaggtt gctgggcact gataattccg tggtgtt
//
