LOCUS       sequence-412579-        6627 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 6627)
  TITLE     Prigge Lab AAV vectors
REFERENCE   2  (bases 1 to 6627)
  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..6627
                     /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..2970
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        3071..4249
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3302..4240
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4197..4217
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(4280..4313)
                     /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(4364..4397)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4409..4435)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(6065..6091)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    6112..6145
                     /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    6196..6229
                     /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(6242..6627,1..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 cgccttagcg 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 acactcaact ctatctcggg
      661 ctattctttt gatttataag ggattttgcc gatttcggtc 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 atactgttct
     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 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     2941 gagtggccaa ctccatcact aggggttcct gcggccgcac gcgtaagctt tgcaaagatg
     3001 gataaagttt taaacagaga ggaatctttg cagctaatgg accttctagg tcttgaaagg
     3061 agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     3121 gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa
     3181 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     3241 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     3301 ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt
     3361 gccttgaatt acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg
     3421 aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt
     3481 tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg
     3541 tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct
     3601 ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt
     3661 ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg
     3721 gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc
     3781 tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg
     3841 gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc
     3901 aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag
     3961 ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag
     4021 gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt
     4081 ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca
     4141 cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa
     4201 gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc
     4261 ctctagagtc gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat
     4321 ggtagctgga ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat
     4381 acattatacg aagttatggc gcgccttaag cgtaatctgg aacatcgtat gggtaaaact
     4441 ccccggaccc agctgctaaa tggctctgat acaagctgtg gtaatcctct ttctccatga
     4501 ggagtggctg cttttcttca ggaagctgct ttctcttgtg acgacacagc aagctcacaa
     4561 gccctgccag cagggcagtg aggacggccc ctaccatcgc cgccccaagg agccatgacc
     4621 agatccgact cgcttgttcc aaataggact taatgtagtc ttgaaaagag tctgggtctg
     4681 aatcttgtag atagctatag tcatagccca gatctttgga tgaaataaag aaatcaccat
     4741 ttctgtacag tggtataaaa ggaaccatgt aggattcccg gttatgtcca atgggtgcat
     4801 tggcttctgg ataaacttct tgaagaggac ggtgccttcg gagccactgc tcaaaaatac
     4861 tgtcaacaaa tgcatggtga agaaggaaga taggatcgtt ggcagatccc tgtacctggg
     4921 acattgttcc attcatatag atgtgcaagg cattgtgcat gctgctttga gaggcatccg
     4981 ctatcccagt aagtggacta gcaaatcctt ccagtgtatt tctaaagctg aaattggcag
     5041 ctttatccat ggaaccagat tcatattggg tcaaactcag gcaaaattct acatcagctg
     5101 aagaggggag ccttggggtt ctggatttgt catggtttcc aggattacgc cgtaaaggtc
     5161 cctcgggcgt tccattgcat aaagactgat ggctgttgta ctcctccaat cggctacaga
     5221 caatctgcca agaggagaag aatgatgctg ggctgagtaa gttaggattt gtggggtgct
     5281 gacctcccat gtactcatct gtgcaaatgt cacacttttc tgcatcccgc cagtcccaat
     5341 atggaatagt gaagttttca tctcctgtca gcttctggat ttcttgttcc caccgcaaca
     5401 agaagagtct atgccaaggc agaaaagctg gtgcttcatg ggcaaaatca atgtctctcc
     5461 agatttcaga tcccccaagc agtgcatcca ttgacacata ataatgttac atccagacaa
     5521 agaggtcata aatattgatg tcgttaaaca tgggtgttga tccatttttc atttggccat
     5581 aggtccctat ggggatgaca tagtctgagc tgatggtatg ctttgctaaa gtgaggtagg
     5641 caaaaaattt gtccttctct ggggcactca aatcgaagat gtttcttctc accaagagtc
     5701 gtctctctgt gcagtttggt ccccaaaagc caaacttgca gtttccacag ttgaatccca
     5761 tgaagttgcc agagcactgg caggtcctat tataaaagac ggaaggccac gactcccggt
     5821 catccacccc tgtgaaggga aattgaggcc caagtggtgc attggacaga aggatattct
     5881 gacaggaacc tctgcctgaa agctggccac agggactcct gtccccgctc cacggtggac
     5941 agcattcctt ctccatcagg ttcttagagg agacacaggc tctagggaaa tggccagcgg
     6001 aggtctggaa actccacagc aggcagtaca aaacagccag gagaaactcc ccggacccag
     6061 ctgcagcgta atctggaaca tcgtatgggt acatcaaggt cttaggctag cataacttcg
     6121 tataaagtat cctatacgaa gttatttgcc ttaacccaga aattatcact gttattcttt
     6181 agaatggtgc aaagaataac ttcgtataat gtatgctata cgaagttatg aattcaagct
     6241 taatcaacct ctggattaca aaatttgtga aagattgact gatattctta actatgttgc
     6301 tccttttacg ctgtgtggat atgctgcttt aatgcctctg tatcatgcta ttgcttcccg
     6361 tatggctttc gttttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     6421 gtggcccgtt gtccgtcaac gtggcgtggt gtgctctgtg tttgctgacg caacccccac
     6481 tggctggggc attgccacca cctgtcaact cctttctggg actttcgctt tccccctccc
     6541 gatcgccacg gcagaactca tcgccgcctg ccttgcccgc tgctggacag gggctaggtt
     6601 gctgggcact gataattccg tggtgtt
//
