LOCUS       sequence-361419-        5097 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To produce AAV with the P3 promoter to express IgkL-mNeonGreen (IgKL
            is a secretory signal peptide).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5097)
  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 5097)
  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-IgKL-mNG
FEATURES             Location/Qualifiers
     source          1..5097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     CDS             576..1283
                     /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      828..837
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1310..1898
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1363..1383)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1781..1792)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1916..2140
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2185..2325
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2185..2314
                     /label=AAV2 ITR
     rep_origin      2400..2855
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2487..2506)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2697..2718
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2872..2891)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2991..3013
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3051..3069)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3137..3241
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3242..3310,3311..4102)
                     /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(3460..3479)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4273..4861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4762..4781
                     /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 ccaccatgac agacacactc ctgctatggg
      541 tactgctgct ctgggttcca ggttccactg gtgacgtgag caagggcgag gaggataaca
      601 tggcctctct cccagcgaca catgagttac acatctttgg ctccatcaac ggtgtggact
      661 ttgacatggt gggtcagggc accggcaatc caaatgatgg ttatgaggag ttaaacctga
      721 agtccaccaa gggtgacctc cagttctccc cctggattct ggtccctcat atcgggtatg
      781 gcttccatca gtacctgccc taccctgacg ggatgtcgcc tttccaggcc gccatggtag
      841 atggctccgg ctaccaagtc catcgcacaa tgcagtttga agatggtgcc tcccttactg
      901 ttaactaccg ctacacctac gagggaagcc acatcaaagg agaggcccag gtgaagggga
      961 ctggtttccc tgctgacggt cctgtgatga ccaactcgct gaccgctgcg gactggtgca
     1021 ggtcgaagaa gacttacccc aacgacaaaa ccatcatcag tacctttaag tggagttaca
     1081 ccactggaaa tggcaagcgc taccggagca ctgcgcggac cacctacacc tttgccaagc
     1141 caatggcggc taactatctg aagaaccagc cgatgtacgt gttccgtaag acggagctca
     1201 agcactccaa gaccgagctc aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg
     1261 gcatggacga gctgtacaag taaagctaag cggccgcaag cttatcgata atcaacctct
     1321 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     1381 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     1441 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     1501 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     1561 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     1621 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     1681 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     1741 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     1801 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     1861 tcagacgagt cggatctccc tttgggccgc ctccccgctc tcgagatacg tacgactgtg
     1921 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1981 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2041 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2101 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagatct
     2161 accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     2221 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     2281 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     2341 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     2401 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2461 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2521 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2581 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     2641 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2701 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     2761 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2821 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     2881 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     2941 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     3001 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     3061 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     3121 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3181 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3241 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3301 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3361 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3421 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3481 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3541 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3601 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3661 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3721 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3781 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3841 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3901 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag
     3961 ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4021 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4081 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     4141 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4201 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4261 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4321 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4381 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4441 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4501 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4561 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4621 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4681 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4741 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4801 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4861 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4921 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4981 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     5041 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcag
//
