LOCUS       sequence-361420-        6911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To produce AAV with the P3 promoter to express Alb(mouse)-mScarlet 
            fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6911)
  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 6911)
  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-mScarlet
FEATURES             Location/Qualifiers
     source          1..6911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     CDS             2397..3089
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(2527..2545)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2795..2814
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3124..3712
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3177..3197)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3595..3606)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3730..3954
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3999..4139
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3999..4128
                     /label=AAV2 ITR
     rep_origin      4214..4669
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4301..4320)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4511..4532
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4686..4705)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4805..4827
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4865..4883)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4951..5055
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5056..5124,5125..5916)
                     /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(5274..5293)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6087..6675
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6576..6595
                     /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 ggcagtgatc aaggagttca tgcggttcaa ggtgcacatg gagggctcca
     2461 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     2521 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttctcctgg gacatcctgt
     2581 cccctcagtt catgtacggc tccagggcct tcaccaagca ccccgccgac atccccgact
     2641 actataagca gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     2701 gcggcgccgt gaccgtgacc caggacacct ccctggagga cggcaccctg atctacaagg
     2761 tgaagctccg cggcaccaac ttccctcctg acggccccgt aatgcagaag aagacaatgg
     2821 gctgggaagc gtccaccgag cggttgtacc ccgaggacgg cgtgctgaag ggcgacatta
     2881 agatggccct gcgcctgaag gacggtggcc gctacctggc ggacttcaag accacctaca
     2941 aggccaagaa gcccgtgcag atgcccggcg cctacaacgt cgaccgcaag ttggacatca
     3001 cctcccacaa cgaggactac accgtggtgg aacagtacga acgctccgag ggccgccact
     3061 ccaccggcgg catggacgag ctgtacaagt agcggccgcg aattcgatat caagcttatc
     3121 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3181 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3241 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3301 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3361 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3421 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3481 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     3541 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     3601 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     3661 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gctctcgaga
     3721 tacgtacgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3781 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3841 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3901 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3961 aggcggaaag atctaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     4021 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4081 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     4141 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     4201 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4261 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     4321 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4381 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     4441 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4501 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     4561 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     4621 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     4681 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     4741 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4801 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     4861 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4921 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4981 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5041 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5101 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5161 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5221 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5281 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5341 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5401 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5461 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5521 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5581 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5641 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5701 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5761 ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5821 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5881 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5941 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6001 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6061 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6121 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6181 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6241 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6301 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6361 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6421 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6481 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6541 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6601 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6661 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6721 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6781 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6841 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6901 cattaatgca g
//
