LOCUS       sequence-471028-        6906 bp    DNA     circular SYN 30-APR-2025
DEFINITION  To produce AAV with the P3 promoter to express 
            Alb(mouse)-mStayGold(Ando) fusion protein .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6906)
  TITLE     Hirase lab plasmids
REFERENCE   2  (bases 1 to 6906)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAAV-P3-Alb-mStayGold
FEATURES             Location/Qualifiers
     source          1..6906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..675
                     /codon_start=1
                     /product="bright and highly photostable monomeric green 
                     fluorescent protein (Ando et al., 2023)"
                     /label=mStayGold(J)
                     /note="includes multiple mutations plus an N-terminal 
                     iinker sequence"
                     /translation="VSTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKGK
                     YVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGSI
                     HAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMDVSLPNEVQHIPIDDGVECTVTLQ
                     YPLLSDESKCVEAYQNTIIKPLHNQPAPDVPFHWIRKQYTQSKDDTEERDHIIQSETLE
                     AHL"
     misc_feature    723..1311
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(776..796)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1194..1205)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1329..1553
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1598..1738
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1598..1727
                     /label=AAV2 ITR
     rep_origin      1813..2268
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1900..1919)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2110..2131
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2285..2304)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2404..2426
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2464..2482)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2550..2654
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2655..2723,2724..3515)
                     /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(2873..2892)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3686..4274
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4175..4194
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4511..4640
                     /label=AAV2 ITR
ORIGIN
        1 gtgtctacag gcgaggagct gtttaccggc gtggtgccct tcaagttcca gctgaagggc
       61 accatcaacg gcaagagctt caccgtggaa ggcgagggcg agggcaatag ccacgagggc
      121 agccacaaag gcaagtacgt gtgcaccagc ggcaaactgc caatgtcttg ggccgccctg
      181 ggaactagct tcggctatgg catgaagtac tacaccaagt accccagcgg cctgaagaac
      241 tggttccacg aggtgatgcc cgagggcttc acctacgaca gacacatcca gtacaagggc
      301 gacggcagca tccacgccaa gcaccagcac ttcatgaaga acggcaccta ccacaacatc
      361 gtggagttca ccggccagga cttcaaggag aacagccccg tgctgaccgg cgacatggac
      421 gtgagcctgc ccaacgaggt gcagcacatc cccatagatg acggcgtgga gtgcacagtg
      481 accctgcagt accctctgct gagcgacgag agcaagtgcg tggaggccta ccagaacacc
      541 atcatcaagc ccctgcacaa tcagccagcc cccgatgtgc cattccactg gatcagaaag
      601 cagtacaccc agagcaagga cgacaccgag gagagagacc acatcatcca gagcgagacc
      661 ctggaggccc acctgtaagc ggccgcaagc tttctagaga attcgatatc aagcttatcg
      721 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
      781 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
      841 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
      901 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
      961 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     1021 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     1081 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     1141 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     1201 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     1261 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg ctctcgagat
     1321 acgtacgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     1381 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     1441 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     1501 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     1561 ggcggaaaga tctaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     1621 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     1681 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     1741 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     1801 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1861 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     1921 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     1981 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     2041 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2101 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     2161 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     2221 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     2281 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     2341 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2401 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     2461 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     2521 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2581 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2641 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2701 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2761 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2821 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2881 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2941 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3001 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3061 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3121 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3181 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3241 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3301 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3361 gtgagcgtgg aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3421 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3481 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3541 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3601 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3661 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3721 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3781 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3841 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3901 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3961 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4021 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4081 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4141 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4201 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4261 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4321 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4381 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4441 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4501 attaatgcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg
     4561 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
     4621 ctccatcact aggggttcct tgtagttaat gattaacccg ccatgctact tatctactcg
     4681 agaccgggcg cgccggggga ggctgctggt gaatattaac caaggtcacc ccagttatcg
     4741 gaggagcaaa caggggctaa gtccacacgc gtggtaccgt ctgtctgcac atttcgtaga
     4801 gcgagtgttc cgatactcta atctccctag gcaaggttca tatttgtgta ggttacttat
     4861 tctccttttg ttgactaagt caataatcag aatcagcagg tttggagtca gcttggcagg
     4921 gatcagcagc ctgggttgga aggagggggt ataaaagccc cttcaccagg agaagccgtc
     4981 acacagatcc acaagctcct ggtcgacact agtaccggtg ccaccatgaa gtgggtaacc
     5041 tttctcctcc tcctcttcgt ctccggctct gctttttcca ggggtgtgtt tcgccgagaa
     5101 gcacacaaga gtgagatcgc ccatcggtat aatgatttgg gagaacaaca tttcaaaggc
     5161 ctagtcctga ttgccttttc ccagtatctc cagaaatgct catacgatga gcatgccaaa
     5221 ttagtgcagg aagtaacaga ctttgcaaag acgtgtgttg ccgatgagtc tgccgccaac
     5281 tgtgacaaat cccttcacac tctttttgga gataagttgt gtgccattcc aaacctccgt
     5341 gaaaactatg gtgaactggc tgactgctgt acaaaacaag agcccgaaag aaacgaatgt
     5401 ttcctgcaac acaaagatga caaccccagc ctgccaccat ttgaaaggcc agaggctgag
     5461 gccatgtgca cctcctttaa ggaaaaccca accaccttta tgggacacta tttgcatgaa
     5521 gttgccagaa gacatcctta tttctatgcc ccagaacttc tttactatgc tgagcagtac
     5581 aatgagattc tgacccagtg ttgtgcagag gctgacaagg aaagctgcct gaccccgaag
     5641 cttgatggtg tgaaggagaa agcattggtc tcatctgtcc gtcagagaat gaagtgctcc
     5701 agtatgcaga agtttggaga gagagctttt aaagcatggg cagtagctcg tctgagccag
     5761 acattcccca atgctgactt tgcagaaatc accaaattgg caacagacct gaccaaagtc
     5821 aacaaggagt gctgccatgg tgacctgctg gaatgcgcag atgacagggc ggaacttgcc
     5881 aagtacatgt gtgaaaacca ggcgactatc tccagcaaac tgcagacttg ctgcgataaa
     5941 ccactgttga agaaagccca ctgtcttagt gaggtggagc atgacaccat gcctgctgat
     6001 ctgcctgcca ttgctgctga ttttgttgag gaccaggaag tgtgcaagaa ctatgctgag
     6061 gccaaggatg tcttcctggg cacgttcttg tatgaatatt caagaagaca ccctgattac
     6121 tctgtatccc tgttgctgag acttgctaag aaatatgaag ccactctgga aaagtgctgc
     6181 gctgaagcca atcctcccgc atgctacggc acagtgcttg ctgaatttca gcctcttgta
     6241 gaagagccta agaacttggt caaaaccaac tgtgatcttt acgagaagct tggagaatat
     6301 ggattccaaa atgccattct agttcgctac acccagaaag cacctcaggt gtcaacccca
     6361 actctcgtgg aggctgcaag aaacctagga agagtgggca ccaagtgttg tacacttcct
     6421 gaagatcaga gactgccttg tgtggaagac tatctgtctg caatcctgaa ccgtgtgtgt
     6481 ctgctgcatg agaagacccc agtgagtgag catgttacca agtgctgtag tggatccctg
     6541 gtggaaaggc ggccatgctt ctctgctctg acagttgatg aaacatatgt ccccaaagag
     6601 tttaaagctg agaccttcac cttccactct gatatctgca cacttccaga gaaggagaag
     6661 cagattaaga aacaaacggc tcttgctgag ctggtgaagc acaagcccaa ggctacagcg
     6721 gagcaactga agactgtcat ggatgacttt gcacagttcc tggatacatg ttgcaaggct
     6781 gctgacaagg acacctgctt ctcgactgag ggtccaaacc ttgtcactag atgcaaagac
     6841 gccttagccc ccgggggtgg aggcggttca ggcggaggtg gctctggcgg tggcggatca
     6901 gtcgac
//
