LOCUS       sequence-361425-        6936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To produce AAV with the P3 promoter to express Alb(mouse)-mNeonGreen
            fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6936)
  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 6936)
  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-mNG
FEATURES             Location/Qualifiers
     source          1..6936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     CDS             2397..3104
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTDVMGMDELYK"
     regulatory      2649..2658
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3149..3737
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3202..3222)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3620..3631)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3755..3979
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4024..4164
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4024..4153
                     /label=AAV2 ITR
     rep_origin      4239..4694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4326..4345)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4536..4557
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4711..4730)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4830..4852
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4890..4908)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4976..5080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5081..5149,5150..5941)
                     /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(5299..5318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6112..6700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6601..6620
                     /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 ggaggataac atggcctctc tcccagcgac acatgagtta cacatctttg
     2461 gctccatcaa cggtgtggac tttgacatgg tgggtcaggg caccggcaat ccaaatgatg
     2521 gttatgagga gttaaacctg aagtccacca agggtgacct ccagttctcc ccctggattc
     2581 tggtccctca tatcgggtat ggcttccatc agtacctgcc ctaccctgac gggatgtcgc
     2641 ctttccaggc cgccatggta gatggctccg gataccaagt ccatcgcaca atgcagtttg
     2701 aagatggtgc ctcccttact gttaactacc gctacaccta cgagggaagc cacatcaaag
     2761 gagaggccca ggtgaagggg actggtttcc ctgctgacgg tcctgtgatg accaactcgc
     2821 tgaccgctgc ggactggtgc aggtcgaaga agacttaccc caacgacaaa accatcatca
     2881 gtacctttaa gtggagttac accactggaa atggcaagcg ctaccggagc actgcgcgga
     2941 ccacctacac ctttgccaag ccaatggcgg ctaactatct gaagaaccag ccgatgtacg
     3001 tgttccgtaa gacggagctc aagcactcca agaccgagct caacttcaag gagtggcaaa
     3061 aggcctttac cgatgtgatg ggcatggacg agctgtacaa gtaagcggcc gcaagctttc
     3121 tagagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     3181 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3241 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3301 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3361 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3421 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3481 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3541 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3601 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3661 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3721 ttgggccgcc tccccgctct cgagatacgt acgactgtgc cttctagttg ccagccatct
     3781 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3841 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3901 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3961 gatgcggtgg gctctatggc ttctgaggcg gaaagatcta ccacgtgcgg accgagcggc
     4021 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4081 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4141 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     4201 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4261 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     4321 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4381 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4441 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4501 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4561 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     4621 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     4681 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4741 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4801 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4861 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4921 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4981 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5041 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5101 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5161 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5221 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5281 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5341 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5401 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5461 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5521 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5581 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5641 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5701 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5761 gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca ttgcagcact
     5821 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5881 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5941 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6001 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6061 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6121 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6181 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6241 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6301 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6361 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6421 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6481 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6541 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6601 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6661 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6721 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6781 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6841 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     6901 gcctctcccc gcgcgttggc cgattcatta atgcag
//
