LOCUS       sequence-423990-        6866 bp    DNA     circular SYN 15-APR-2025
DEFINITION  To produce AAV with the P3 promoter to express Alb(mouse)-StayGold 
            fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6866)
  TITLE     AAV plasmid to produce Albumin-StayGold fusion protein to label 
            blood plasma via liver hepatocyte secretion.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6866)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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            "Unpublished"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-P3-Alb-StayGold
FEATURES             Location/Qualifiers
     source          1..6866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     CDS             2397..3044
                     /codon_start=1
                     /product="bright and highly photostable dimeric green 
                     fluorescent protein (Hirano et al., 2022)"
                     /label=StayGold
                     /note="engineered variant of a GFP from Cytaeis uchidae"
                     /translation="ASTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKGKYVCTSGKLP
                     MSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGSIHAKHQHFMK
                     NGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVQHIPRDDGVECPVTLLYPLLSDKSK
                     CVEAHQNTICKPLHNQPAPDVPYHWIRKQYTQSKDDTEERDHICQSETLEAHL"
     misc_feature    3079..3667
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3132..3152)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3550..3561)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3685..3909
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3954..4094
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3954..4083
                     /label=AAV2 ITR
     rep_origin      4169..4624
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4256..4275)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4466..4487
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4641..4660)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4760..4782
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4820..4838)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4906..5010
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5011..5079,5080..5871)
                     /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(5229..5248)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6042..6630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6531..6550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctactcg agaccgggcg
      181 cgccggggga ggctgctggt gaatattaac caaggtcacc ccagttatcg gaggagcaaa
      241 caggggctaa gtccacacgc gtggtaccgt ctgtctgcac atttcgtaga gcgagtgttc
      301 cgatactcta atctccctag gcaaggttca tatttgtgta ggttacttat tctccttttg
      361 ttgactaagt caataatcag aatcagcagg tttggagtca gcttggcagg gatcagcagc
      421 ctgggttgga aggagggggt ataaaagccc cttcaccagg agaagccgtc acacagatcc
      481 acaagctcct ggtcgacact agtaccggtg ccaccatgaa gtgggtaacc tttctcctcc
      541 tcctcttcgt ctccggctct gctttttcca ggggtgtgtt tcgccgagaa gcacacaaga
      601 gtgagatcgc ccatcggtat aatgatttgg gagaacaaca tttcaaaggc ctagtcctga
      661 ttgccttttc ccagtatctc cagaaatgct catacgatga gcatgccaaa ttagtgcagg
      721 aagtaacaga ctttgcaaag acgtgtgttg ccgatgagtc tgccgccaac tgtgacaaat
      781 cccttcacac tctttttgga gataagttgt gtgccattcc aaacctccgt gaaaactatg
      841 gtgaactggc tgactgctgt acaaaacaag agcccgaaag aaacgaatgt ttcctgcaac
      901 acaaagatga caaccccagc ctgccaccat ttgaaaggcc agaggctgag gccatgtgca
      961 cctcctttaa ggaaaaccca accaccttta tgggacacta tttgcatgaa gttgccagaa
     1021 gacatcctta tttctatgcc ccagaacttc tttactatgc tgagcagtac aatgagattc
     1081 tgacccagtg ttgtgcagag gctgacaagg aaagctgcct gaccccgaag cttgatggtg
     1141 tgaaggagaa agcattggtc tcatctgtcc gtcagagaat gaagtgctcc agtatgcaga
     1201 agtttggaga gagagctttt aaagcatggg cagtagctcg tctgagccag acattcccca
     1261 atgctgactt tgcagaaatc accaaattgg caacagacct gaccaaagtc aacaaggagt
     1321 gctgccatgg tgacctgctg gaatgcgcag atgacagggc ggaacttgcc aagtacatgt
     1381 gtgaaaacca ggcgactatc tccagcaaac tgcagacttg ctgcgataaa ccactgttga
     1441 agaaagccca ctgtcttagt gaggtggagc atgacaccat gcctgctgat ctgcctgcca
     1501 ttgctgctga ttttgttgag gaccaggaag tgtgcaagaa ctatgctgag gccaaggatg
     1561 tcttcctggg cacgttcttg tatgaatatt caagaagaca ccctgattac tctgtatccc
     1621 tgttgctgag acttgctaag aaatatgaag ccactctgga aaagtgctgc gctgaagcca
     1681 atcctcccgc atgctacggc acagtgcttg ctgaatttca gcctcttgta gaagagccta
     1741 agaacttggt caaaaccaac tgtgatcttt acgagaagct tggagaatat ggattccaaa
     1801 atgccattct agttcgctac acccagaaag cacctcaggt gtcaacccca actctcgtgg
     1861 aggctgcaag aaacctagga agagtgggca ccaagtgttg tacacttcct gaagatcaga
     1921 gactgccttg tgtggaagac tatctgtctg caatcctgaa ccgtgtgtgt ctgctgcatg
     1981 agaagacccc agtgagtgag catgttacca agtgctgtag tggatccctg gtggaaaggc
     2041 ggccatgctt ctctgctctg acagttgatg aaacatatgt ccccaaagag tttaaagctg
     2101 agaccttcac cttccactct gatatctgca cacttccaga gaaggagaag cagattaaga
     2161 aacaaacggc tcttgctgag ctggtgaagc acaagcccaa ggctacagcg gagcaactga
     2221 agactgtcat ggatgacttt gcacagttcc tggatacatg ttgcaaggct gctgacaagg
     2281 acacctgctt ctcgactgag ggtccaaacc ttgtcactag atgcaaagac gccttagccc
     2341 ccgggggtgg aggcggttca ggcggaggtg gctctggcgg tggcggatca gtcgacgcat
     2401 ctactccctt caagttccaa ttgaagggta ctatcaatgg gaagtccttc acagtagagg
     2461 gagagggaga gggaaatagt cacgagggtt cccacaaggg gaagtacgtt tgtacttccg
     2521 gaaagttgcc catgtcctgg gctgcactcg ggacatcctt cgggtacggt atgaagtact
     2581 acactaagta cccatcaggt ctcaagaatt ggttccacga ggttatgcca gagggattca
     2641 cgtacgacag acacatccaa tacaagggtg acggatccat ccacgcaaag caccaacact
     2701 tcatgaagaa tggtacttac cacaatatcg tcgagttcac tgggcaagac ttcaaggaga
     2761 actctcccgt actcaccggg gacatgaatg tctccctacc aaatgaggta caacacatcc
     2821 ctagggacga cggggttgag tgtccagtca cactattgta ccccctactc tccgacaaga
     2881 gtaagtgtgt agaggcccac caaaatacaa tatgtaagcc ccttcacaat caaccagccc
     2941 cagacgtccc ataccactgg atcaggaagc aatacactca atccaaggac gacactgagg
     3001 agcgggacca catctgtcaa tcagagacac tcgaggccca cctttagcgg ccgcgaattc
     3061 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3121 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3181 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3241 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3301 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3361 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3421 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3481 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3541 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3601 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3661 tccccgctct cgagatacgt acgactgtgc cttctagttg ccagccatct gttgtttgcc
     3721 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3781 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3841 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3901 gctctatggc ttctgaggcg gaaagatcta ccacgtgcgg accgagcggc cgcaggaacc
     3961 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4021 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4081 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4141 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4201 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     4261 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4321 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4381 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4441 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4501 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     4561 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4621 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4681 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4741 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4801 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4861 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4921 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4981 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5041 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5101 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5161 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5221 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5281 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5341 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5401 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5461 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5521 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5581 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5641 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5701 gataaatctg gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat
     5761 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5821 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5881 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5941 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6001 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6061 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6121 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6181 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6241 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6301 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6361 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6421 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6481 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6541 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6601 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6661 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6721 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6781 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6841 gcgcgttggc cgattcatta atgcag
//
