LOCUS       sequence-361424-        8570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To produce AAV with the CBh promoter to express 
            Alb(mouse)-mNeonGreen fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8570)
  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 8570)
  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-CBh-Alb-mNG
FEATURES             Location/Qualifiers
     source          1..8570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        124..409
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        411..685
                     /label=chicken beta-actin promoter
     intron          686..913
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             2812..3519
                     /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      3064..3073
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3534..4122
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3587..3607)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4005..4016)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4249..4643
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     repeat_region   4927..5056
                     /label=AAV2 ITR
     rep_origin      5427..5940
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5514..5533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5724..5745
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6422..6441)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6541..6563
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6601..6619)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6687..6791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6792..6860,6861..7652)
                     /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(7010..7029)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7823..8411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8312..8331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttccttg
       61 tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag actcgagacg
      121 cgtcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      181 attgacgtca atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      241 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      301 tgacgtcaat gacggtaaat ggcccgcctg gcattgtgcc cagtacatga ccttatggga
      361 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
      421 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
      481 atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggc gcgcgccagg
      541 cggggcgggg cggggsgrgg ggsggggsgg ggsgrggcgg rgaggtgcgg cggcagccaa
      601 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
      661 taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg
      721 ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
      781 gagcgggcgg gacggccctt ctcctccggg ctgtaattag ctgagcaaga ggtaagggtt
      841 taagggatgg ttggttggtg gggtattaat gtttaattac ctggagcacc tgcctgaaat
      901 cacttttttt caggttggac cggtgccacc atgaagtggg taacctttct cctcctcctc
      961 ttcgtctccg gctctgcttt ttccaggggt gtgtttcgcc gagaagcaca caagagtgag
     1021 atcgcccatc ggtataatga tttgggagaa caacatttca aaggcctagt cctgattgcc
     1081 ttttcccagt atctccagaa atgctcatac gatgagcatg ccaaattagt gcaggaagta
     1141 acagactttg caaagacgtg tgttgccgat gagtctgccg ccaactgtga caaatccctt
     1201 cacactcttt ttggagataa gttgtgtgcc attccaaacc tccgtgaaaa ctatggtgaa
     1261 ctggctgact gctgtacaaa acaagagccc gaaagaaacg aatgtttcct gcaacacaaa
     1321 gatgacaacc ccagcctgcc accatttgaa aggccagagg ctgaggccat gtgcacctcc
     1381 tttaaggaaa acccaaccac ctttatggga cactatttgc atgaagttgc cagaagacat
     1441 ccttatttct atgccccaga acttctttac tatgctgagc agtacaatga gattctgacc
     1501 cagtgttgtg cagaggctga caaggaaagc tgcctgaccc cgaagcttga tggtgtgaag
     1561 gagaaagcat tggtctcatc tgtccgtcag agaatgaagt gctccagtat gcagaagttt
     1621 ggagagagag cttttaaagc atgggcagta gctcgtctga gccagacatt ccccaatgct
     1681 gactttgcag aaatcaccaa attggcaaca gacctgacca aagtcaacaa ggagtgctgc
     1741 catggtgacc tgctggaatg cgcagatgac agggcggaac ttgccaagta catgtgtgaa
     1801 aaccaggcga ctatctccag caaactgcag acttgctgcg ataaaccact gttgaagaaa
     1861 gcccactgtc ttagtgaggt ggagcatgac accatgcctg ctgatctgcc tgccattgct
     1921 gctgattttg ttgaggacca ggaagtgtgc aagaactatg ctgaggccaa ggatgtcttc
     1981 ctgggcacgt tcttgtatga atattcaaga agacaccctg attactctgt atccctgttg
     2041 ctgagacttg ctaagaaata tgaagccact ctggaaaagt gctgcgctga agccaatcct
     2101 cccgcatgct acggcacagt gcttgctgaa tttcagcctc ttgtagaaga gcctaagaac
     2161 ttggtcaaaa ccaactgtga tctttacgag aagcttggag aatatggatt ccaaaatgcc
     2221 attctagttc gctacaccca gaaagcacct caggtgtcaa ccccaactct cgtggaggct
     2281 gcaagaaacc taggaagagt gggcaccaag tgttgtacac ttcctgaaga tcagagactg
     2341 ccttgtgtgg aagactatct gtctgcaatc ctgaaccgtg tgtgtctgct gcatgagaag
     2401 accccagtga gtgagcatgt taccaagtgc tgtagtggat ccctggtgga aaggcggcca
     2461 tgcttctctg ctctgacagt tgatgaaaca tatgtcccca aagagtttaa agctgagacc
     2521 ttcaccttcc actctgatat ctgcacactt ccagagaagg agaagcagat taagaaacaa
     2581 acggctcttg ctgagctggt gaagcacaag cccaaggcta cagcggagca actgaagact
     2641 gtcatggatg actttgcaca gttcctggat acatgttgca aggctgctga caaggacacc
     2701 tgcttctcga ctgagggtcc aaaccttgtc actagatgca aagacgcctt agcccccggg
     2761 ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcagtcga cgtgagcaag
     2821 ggcgaggagg ataacatggc ctctctccca gcgacacatg agttacacat ctttggctcc
     2881 atcaacggtg tggactttga catggtgggt cagggcaccg gcaatccaaa tgatggttat
     2941 gaggagttaa acctgaagtc caccaagggt gacctccagt tctccccctg gattctggtc
     3001 cctcatatcg ggtatggctt ccatcagtac ctgccctacc ctgacgggat gtcgcctttc
     3061 caggccgcca tggtagatgg ctccggatac caagtccatc gcacaatgca gtttgaagat
     3121 ggtgcctccc ttactgttaa ctaccgctac acctacgagg gaagccacat caaaggagag
     3181 gcccaggtga aggggactgg tttccctgct gacggtcctg tgatgaccaa ctcgctgacc
     3241 gctgcggact ggtgcaggtc gaagaagact taccccaacg acaaaaccat catcagtacc
     3301 tttaagtgga gttacaccac tggaaatggc aagcgctacc ggagcactgc gcggaccacc
     3361 tacacctttg ccaagccaat ggcggctaac tatctgaaga accagccgat gtacgtgttc
     3421 cgtaagacgg agctcaagca ctccaagacc gagctcaact tcaaggagtg gcaaaaggcc
     3481 tttaccgatg tgatgggcat ggacgagctg tacaagtaag cggccgcatc gataatcaac
     3541 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3601 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3661 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     3721 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     3781 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     3841 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     3901 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     3961 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     4021 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     4081 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatgc cgtatacggt
     4141 acccggggat cctctagtag agtcgaggaa ttcaccccac cagtgcaggc tgcctatcag
     4201 aaagtggtgg ctggtgtggc taatgccctg gcccacaagt atcactaagc tcgctttctt
     4261 gctgtccaat ttctattaaa ggttcctttg ttccctaagt ccaactacta aactggggga
     4321 tattatgaag ggccttgagc atctggattc tgcctaataa aaaacattta ttttcattgc
     4381 aatgatgtat ttaaattatt tctgaatatt ttactaaaaa gggaatgtgg gaggtcagtg
     4441 catttaaaac ataaagaaat gaagagctag ttcaaacctt gggaaaatac actatatctt
     4501 aaactccatg aaagaaggtg aggctgcaaa cagctaatgc acattggcaa cagcccctga
     4561 tgcctatgcc ttattcatcc ctcagaaaag gattcaagta gaggcttgat ttggaggtta
     4621 aagttttgct atgctgtatt ttacattact tattgtttta gctgtcctca tgaatgtctt
     4681 ttcactaccc atttgcttat cctgcatctc tcagccttga ctccactcag ttctcttgct
     4741 tagagatacc acctttcccc tgaagtgttc cttccatgtt ttacggcgag atggtttctc
     4801 ctcgcctggc cactcagcct tagttgtctc tgttgtctta tagaggtctg gtcgaccccc
     4861 aacgcgttgg agctctagag catggctacg tagataagta gcatggcggg ttaatcatta
     4921 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4981 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     5041 gcgagcgagc gcgcagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     5101 agttgcgcag cctgaatggc gaatggcgat tccgttgcaa tggctggcgg taatattgtt
     5161 ctggatatta ccagcaaggc cgatagtttg agttcttcta ctcaggcaag tgatgttatt
     5221 actaatcaaa gaagtattgc gacaacggtt aatttgcgtg atggacagac tcttttactc
     5281 ggtggcctca ctgattataa aaacacttct caggattctg gcgtaccgtt cctgtctaaa
     5341 atccctttaa tcggcctcct gtttagctcc cgctctgatt ctaacgagga aagcacgtta
     5401 tacgtgctcg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     5461 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5521 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5581 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5641 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5701 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5761 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5821 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     5881 ttaaatattt gcttatacaa tcttcctgtt tttggggctt ttctgattat caaccggggt
     5941 acatatgatt gacatgctag ttttacgatt accgttcatc gattctcttg tttgctccag
     6001 actctcaggc aatgacctga tagcctttgt agagacctct caaaaatagc taccctctcc
     6061 ggcatgaatt tatcagctag aacggttgaa tatcatattg atggtgattt gactgtctcc
     6121 ggcctttctc acccgtttga atctttacct acacattact caggcattgc atttaaaata
     6181 tatgagggtt ctaaaaattt ttatccttgc gttgaaataa aggcttctcc cgcaaaagta
     6241 ttacagggtc ataatgtttt tggtacaacc gatttagctt tatgctctga ggctttattg
     6301 cttaattttg ctaattcttt gccttgcctg tatgatttat tggatgttgg aatcgcctga
     6361 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     6421 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     6481 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     6541 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     6601 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     6661 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6721 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6781 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6841 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6901 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6961 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     7021 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     7081 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     7141 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     7201 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     7261 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     7321 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     7381 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     7441 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7501 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7561 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7621 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     7681 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7741 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7801 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7861 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7921 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7981 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     8041 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     8101 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     8161 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     8221 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     8281 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     8341 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     8401 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     8461 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     8521 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
//
