LOCUS       sequence-237070-        6194 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6194)
  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 6194)
  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..6194
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     repeat_region   417..546
                     /label=AAV2 ITR
     rep_origin      801..1256
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(888..907)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1098..1119
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1282..1386
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1387..1455,1456..2247)
                     /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(1605..1624)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2418..3006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2907..2926
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3243..3372
                     /label=AAV2 ITR
     polyA_signal    3534..3655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3571..3590)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3625..3644
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3698..4286
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3804..3815
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4213..4233
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4422..5132,5133..5135,5136..5138))
                     /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(4464..4485)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4813..4832
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5072..5093
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      5135..5144
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5157..5534)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(5645..5664)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(5726..5748)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 gcccccatcg ctgcacaaaa taattaaaaa ataaataaat acaaaattgg gggtggggag
       61 gggggggaga tggggagagt gaagcagaac gtggggctca cctcgaccat ggtaatagcg
      121 atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca
      181 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga
      241 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
      301 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatggtact
      361 cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta actacaagga
      421 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
      481 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
      541 gcgcagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
      601 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
      661 tacgcgcagc gtgaccgcta cacttgccgg ctttgcccgg gcggcctcag tgagcgagcg
      721 agcgcgcagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
      781 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
      841 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
      901 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
      961 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1021 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1081 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     1141 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     1201 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     1261 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1321 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1381 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1441 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1501 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1561 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1621 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1681 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1741 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1801 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1861 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1921 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1981 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2041 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2101 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     2161 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     2221 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2281 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2341 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2401 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2461 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2521 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2581 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2641 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2701 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2761 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2821 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2881 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2941 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3001 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3061 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3121 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3181 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3241 agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct
     3301 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
     3361 ctaggggttc cttgtagtta atgattaacc cgccatgcta cttatctacg tagccatgct
     3421 ctaggaagat ccttatcgat tttaccacat ttgtagaggt tttacttgct ttaaaaaacc
     3481 tcccacatct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3541 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3601 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3661 tctgctcgaa gcggccgccc gggtcgacgg tatcgatgcg gggaggcggc ccaaagggag
     3721 atccgactcg tctgagggcg aaggcgaaga cgcggaagag gccgcagagc cggcagcagg
     3781 ccgcgggaag gaaggtccgc tggattgagg gccgaaggga cgtagcagaa ggacgtcccg
     3841 cgcagaatcc aggtggcaac acaggcgagc agccaaggaa aggacgatga tttccccgac
     3901 aacaccacgg aattgtcagt gcccaacagc cgagcccctg tccagcagcg ggcaaggcag
     3961 gcggcgatga gttccgccgt ggcaataggg agggggaaag cgaaagtccc ggaaaggagc
     4021 tgacaggtgg tggcaatgcc ccaaccagtg ggggttgcgt cagcaaacac agtgcacacc
     4081 acgccacgtt gcctgacaac gggccacaac tcctcataaa gagacagcaa ccaggattta
     4141 tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa aggcattaaa
     4201 gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt caatctttca
     4261 caaattttgt aatccagagg ttgattatcg ataagcttga tatcgaattg cattcatttt
     4321 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     4381 tgtggtatgg ctgattatga tctagagtcg cggccgcttt acttgtacag ctcgtccatg
     4441 ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg
     4501 ttggggtctt tgctcagggc ggactggtgg ctcaggtagt ggttgtcggg cagcagcacg
     4561 gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc
     4621 tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt tcttctgctt gtcggccatg
     4681 atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc
     4741 tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc
     4801 acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg
     4861 tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg
     4921 ctgaagcact gcacgccgta gctgaaggtg gtcacgaggg tgggccaggg cacgggcagc
     4981 ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc
     5041 tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc
     5101 accaccccgg tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg tggatcctta
     5161 gcgctggtgt acttggtgat ggccttagta ccctcggaca cggcgtgctt ggccaactcc
     5221 ccaggcagca gcaggcgcac ggccgtctgg atctccctgg aggtgatggt cgagcgcttg
     5281 ttgtaatgcg ccaggcggga agcctcacct gcgatgcgct cgaaaatgtc gttcacaaac
     5341 gaattcatga tgcccatggc cttggacgaa atgccggtgt cagggtggac ctgcttcaga
     5401 accttgtaca catagatgga atagctctcc ttgcggctgc gcttgcgctt cttgccgcct
     5461 ttcttctgcg ccttagtcac cgccttcttg gagccctttt tcggggcggg agcagacttc
     5521 gctggctctg gcatggtggc ggtaccgtcg actgcagaat tcgaagcttg agctcgagat
     5581 ctgagtccgg tagcgctagc accggtggat cccgatccaa ttctttgcca aaatgatgag
     5641 acagcacaat aaccagcacg ttgcccagga gctgtaggaa aaagaagaag gcatgaacat
     5701 ggttagcaga ggctctagag ccgccggtca cacgccagaa gccgaacccc gccctgcccc
     5761 gtcccccccg aaggcagccg tccccccgcg gacagccccg agccgctccc cccgcacaaa
     5821 gggccctccc ggagcccctc aaggctttca cgcagccaca gaaaagaaac aagccgtcat
     5881 taaaccaagc gctaattaca gcccggagga gaagggccgt cccgcccgct cacctgtggg
     5941 agtaacgcgg tcagtcagag ccggggcggg cggcgcgagg cggcggcgga gcggggcacg
     6001 gggcgaaggc agcgcgcagc gactcccgcc cgccgcgcgc ttcgcttttt atagggccgc
     6061 cgccgccgcc gcctcgccat aaaaggaaac tttcggagcg cgccgctctg attggctgcc
     6121 gccgcacctc tccgcctcgc cccgccccgc ccctcgcccc gccccgcccc gcctggcgcg
     6181 cgcccccccc cccc
//
