LOCUS       sequence-237072-        6198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6198)
  AUTHORS   Lien AD, Scanziani M
  TITLE     In vivo Labeling of Constellations of Functionally Identified 
            Neurons for Targeted in vitro Recordings.
  JOURNAL   Front Neural Circuits. 2011;5:16. Epub 2011 Nov 29.
  PUBMED    22144948
REFERENCE   2  (bases 1 to 6198)
  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: "Front 
            Neural Circuits."; date: "2011"; volume: "5"; pages: "16. Epub 2011 
            Nov 29"
COMMENT     Sequence Label: pACAGW-H2B-PAGFP-AAV
FEATURES             Location/Qualifiers
     source          1..6198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1235..1945,1946..1948,1949..1951))
                     /codon_start=1
                     /product="photoactivatable GFP variant (Patterson et al., 
                     2002)"
                     /label=PA-GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1277..1298)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1626..1645
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1885..1906
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1948..1957
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1970..2347)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2458..2477)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2539..2561)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(2840..3117)
                     /label=chicken beta-actin promoter
     repeat_region   3428..3557
                     /label=AAV2 ITR
     rep_origin      3812..4267
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3899..3918)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4109..4130
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4293..4397
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4398..4466,4467..5258)
                     /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(4616..4635)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5429..6017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5918..5937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct tgatatcgaa ttgcattcat tttatgtttc
     1141 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     1201 tggctgatta tgatctagag tcgcggccgc tttacttgta cagctcgtcc atgccgagag
     1261 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     1321 ctttgctcag ggcggactgg tggctcaggt agtggttgtc gggcagcagc acggggccgt
     1381 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt
     1441 tgtggcggat cttgaagttg gccttgatgc cgttcttctg cttgtcggcc atgatataga
     1501 cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     1561 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     1621 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     1681 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc
     1741 actgcacgcc gtagctgaag gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     1801 tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg
     1861 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
     1921 cggtgaacag ctcctcgccc ttgctcacca tggtggcgac cggtggatcc ttagcgctgg
     1981 tgtacttggt gatggcctta gtaccctcgg acacggcgtg cttggccaac tccccaggca
     2041 gcagcaggcg cacggccgtc tggatctccc tggaggtgat ggtcgagcgc ttgttgtaat
     2101 gcgccaggcg ggaagcctca cctgcgatgc gctcgaaaat gtcgttcaca aacgaattca
     2161 tgatgcccat ggccttggac gaaatgccgg tgtcagggtg gacctgcttc agaaccttgt
     2221 acacatagat ggaatagctc tccttgcggc tgcgcttgcg cttcttgccg cctttcttct
     2281 gcgccttagt caccgccttc ttggagccct ttttcggggc gggagcagac ttcgctggct
     2341 ctggcatggt ggcggtaccg tcgactgcag aattcgaagc ttgagctcga gatctgagtc
     2401 cggtagcgct agcaccggtg gatcccgatc caattctttg ccaaaatgat gagacagcac
     2461 aataaccagc acgttgccca ggagctgtag gaaaaagaag aaggcatgaa catggttagc
     2521 agaggctcta gagccgccgg tcacacgcca gaagccgaac cccgccctgc cccgtccccc
     2581 ccgaaggcag ccgtcccccc gcggacagcc ccgagccgct ccccccgcac aaagggccct
     2641 cccggagccc ctcaaggctt tcacgcagcc acagaaaaga aacaagccgt cattaaacca
     2701 agcgctaatt acagcccgga ggagaagggc cgtcccgccc gctcacctgt gggagtaacg
     2761 cggtcagtca gagccggggc gggcggcgcg aggcggcggc ggagcggggc acggggcgaa
     2821 ggcagcgcgc agcgactccc gcccgccgcg cgcttcgctt tttatagggc cgccgccgcc
     2881 gccgcctcgc cataaaagga aactttcgga gcgcgccgct ctgattggct gccgccgcac
     2941 ctctccgcct cgccccgccc cgcccctcgc cccgccccgc cccgcctggc gcgcgccccc
     3001 cccccccccc cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg
     3061 ggggtgggga ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca
     3121 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     3181 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     3241 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     3301 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     3361 tcaatggtac tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt
     3421 aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     3481 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     3541 agcgagcgag cgcgcagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     3601 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     3661 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgccg gctttgcccg ggcggcctca
     3721 gtgagcgagc gagcgcgcag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3781 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     3841 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3901 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3961 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4021 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4081 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4141 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4201 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     4261 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4321 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4381 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4441 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4501 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4561 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4621 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4681 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4741 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4801 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4861 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4921 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4981 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5041 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5101 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5161 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5221 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5281 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5341 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5401 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5461 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5521 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5581 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5641 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5701 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5761 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5821 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5881 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5941 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6001 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6061 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6121 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6181 tgagcgagga agcggaag
//
