LOCUS       sequence-361422-        6960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To produce AAV with the P3 promoter to express Alb-mCarmine fusion 
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6960)
  AUTHORS   Wang X, Delle C, Asiminas A, Akther S, Vittani M, Brogger P, Kusk P,
            Vo CT, Radovanovic T, Konno A, Hirai H, Fukuda M, Weikop P, Goldman 
            SA, Nedergaard M, Hirase H
  TITLE     Liver-secreted fluorescent blood plasma markers enable chronic 
            imaging of the microcirculation.
  JOURNAL   Cell Rep Methods. 2022 Sep 21;2(10):100302. doi: 
            10.1016/j.crmeth.2022.100302. eCollection 2022 Oct 24.
  PUBMED    36313804
REFERENCE   2  (bases 1 to 6960)
  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: "Cell Rep 
            Methods."; date: "2022-09-21"; volume: "2(10):100302. doi"; pages: "
            10.1016/j.crmeth.2022.100302. eCollection 2022 Oct 24"
COMMENT     Sequence Label: pAAV-P3-Alb-mCarmine
FEATURES             Location/Qualifiers
     source          1..6960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     CDS             2397..3128
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein with 
                     favorable expression and pKa properties and with peak 
                     emission at 675 nm (Fabritius et al., 2018)"
                     /label=mCarmine
                     /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTQR
                     IKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LKVKQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTETLYPADGGLRGYNTM
                     ALKLVGGGHLQTFLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADKETYVEQHEVAVA
                     RYCDLPSKLGHKLNGMDELYK"
     misc_feature    3173..3761
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3226..3246)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3644..3655)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3779..4003
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4048..4188
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4048..4177
                     /label=AAV2 ITR
     rep_origin      4263..4718
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4350..4369)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4560..4581
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4735..4754)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4854..4876
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4914..4932)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5000..5104
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5105..5173,5174..5965)
                     /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(5323..5342)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6136..6724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6625..6644
                     /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 gtcgacgtga
     2401 gcaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
     2461 tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
     2521 agacccagag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     2581 ctaccagctt catgtacggc agcaagacct tcatcaacca cacccagggc atccccgatt
     2641 tctttaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
     2701 ggggcgtgct gaaggtgaaa caggacacca gcctccagga cggctgcctc atctacaacg
     2761 tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg
     2821 gctgggaggc caacaccgag accctgtacc ccgctgacgg cggcctgcgc ggctataaca
     2881 cgatggccct gaagctcgtg ggcgggggcc acctgcagac gttcctcaag accacataca
     2941 gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactttgtg gaccgcagac
     3001 tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag gtggctgtgg
     3061 ccagatactg cgacctccct agcaaactgg ggcacaaact taatggcatg gacgagctgt
     3121 acaagtgagc ggccgcaagc tttctagaga attcgatatc aagcttatcg ataatcaacc
     3181 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3241 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3301 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3361 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3421 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3481 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3541 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     3601 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3661 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3721 ccctcagacg agtcggatct ccctttgggc cgcctccccg ctctcgagat acgtacgact
     3781 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3841 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3901 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3961 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     4021 tctaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     4081 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     4141 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     4201 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     4261 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4321 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4381 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4441 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     4501 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4561 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     4621 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     4681 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     4741 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4801 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4861 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4921 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4981 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5041 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5101 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5161 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5221 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5281 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5341 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5401 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5461 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5521 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5581 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5641 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5701 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5761 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5821 aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5881 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5941 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6001 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6061 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6121 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6181 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6241 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6301 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6361 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6421 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6481 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6541 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6601 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6661 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6721 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6781 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6841 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6901 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
//
