LOCUS       sequence-319192-        5155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing iCre AAV Virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5155)
  AUTHORS   Graybuck LT, Daigle TL, Sedeno-Cortes AE, Walker M, Kalmbach B, Lenz
            GH, Morin E, Nguyen TN, Garren E, Bendrick JL, Kim TK, Zhou T, 
            Mortrud M, Yao S, Siverts LA, Larsen R, Gore BB, Szelenyi ER, Trader
            C, Balaram P, van Velthoven CTJ, Chiang M, Mich JK, Dee N, Goldy J, 
            Cetin AH, Smith K, Way SW, Esposito L, Yao Z, Gradinaru V, Sunkin 
            SM, Lein E, Levi BP, Ting JT, Zeng H, Tasic B
  TITLE     Enhancer viruses for combinatorial cell-subclass-specific labeling.
  JOURNAL   Neuron. 2021 Mar 23. pii: S0896-6273(21)00159-8. doi: 
            10.1016/j.neuron.2021.03.011.
  PUBMED    33789083
REFERENCE   2  (bases 1 to 5155)
  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: "Neuron. 
            2021 Mar 23. pii: S0896-6273(21)00159-8. doi: 
            10.1016/j.neuron.2021.03.011."
COMMENT     Sequence Label: CN1294-scAAV-hI56i-minBglobin-iCre-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             824..844
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1960..1980)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2160..2271
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2510..2527)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2510..2526)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2519..2541)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2735..2754)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2854..2876
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2914..2932)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3000..3104
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3105..3173,3174..3965)
                     /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(3323..3342)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4136..4724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4625..4644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5012..5033
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5048..5053,5054..5071,5072..5078)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5086..5102
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5091..5113
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5110..5126
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5110..5126
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaagcttccc ggggggatct gggccactcc ctctctgcgc gctcgctcgc tcactgaggc
       61 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
      121 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctggaggg gtggagtcgt
      181 gacctaggac gcgtataggt accgagctct atgcactcac agtggtttgg catgcatctg
      241 gtgaattttt tttaacgaaa aattagtgtt ggtttcgatg tatggtagca ttctccctaa
      301 cgtaatttga ataattcagc aaagccccac taccagctgt acttctgcag cctcttccat
      361 tcttttcagc attataattt tggttaattt tcaattttag gtcctacgtc tctgcaattt
      421 gtgtatgaat aacagaataa tttccctctt ttgtttcgcc tttcctgttc ctgaatctaa
      481 ataaagatgg ctttttagta ttaaaagtgg aagaaaatta caggtaatta tctttgacgg
      541 taaaaacgct gtaatcagcg ggctacatga aaaattactc taattatggc tgcatttaag
      601 agaatggaaa aaaaccttct tgtggataaa aaccttaaat tgtccccaat gtctgcttca
      661 aattggatgg cactgcagct ggaggctttg ttcagaattg atcctgggga gctacgaacc
      721 caaagtttca cagtagggag ctcgggctgg gcataaaagt cagggcagag ccatctattg
      781 cttacatttg cttctgggat ccttcgaagc tagcaccatg gtgcccaaga agaagaggaa
      841 agtctccaac ctgctgactg tgcaccaaaa cctgcctgcc ctccctgtgg atgccacctc
      901 tgatgaagtc aggaagaacc tgatggacat gttcagggac aggcaggcct tctctgaaca
      961 cacctggaag atgctcctgt ctgtgtgcag atcctgggct gcctggtgca agctgaacaa
     1021 caggaaatgg ttccctgctg aacctgagga tgtgagggac tacctcctgt acctgcaagc
     1081 cagaggcctg gctgtgaaga ccatccaaca gcacctgggc cagctcaaca tgctgcacag
     1141 gagatctggc ctgcctcgcc cttctgactc caatgctgtg tccctggtga tgaggagaat
     1201 cagaaaggag aatgtggatg ctggggagag agccaagcag gccctggcct ttgaacgcac
     1261 tgactttgac caagtcagat ccctgatgga gaactctgac agatgccagg acatcaggaa
     1321 cctggccttc ctgggcattg cctacaacac cctgctgcgc attgccgaaa ttgccagaat
     1381 cagagtgaag gacatctccc gcaccgatgg tgggagaatg ctgatccaca ttggcaggac
     1441 caagaccctg gtgtccacag ctggtgtgga gaaggccctg tccctggggg ttaccaagct
     1501 ggtggagaga tggatctctg tgtctggtgt ggctgatgac cccaacaact acctgttctg
     1561 ccgggtcaga aagaatggtg tggctgcccc ttctgccacc tcccaactgt ccacccgggc
     1621 cctggaaggg atctttgagg ccacccaccg cctgatctat ggtgccaagg atgactctgg
     1681 gcagagatac ctggcctggt ctggccactc tgccagagtg ggtgctgcca gggacatggc
     1741 cagggctggt gtgtccatcc ctgaaatcat gcaggctggt ggctggacca atgtgaacat
     1801 tgtgatgaac tacatcagaa acctggactc tgagactggg gccatggtga ggctgctcga
     1861 ggatggggac taatgaggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga
     1921 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     1981 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2041 ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct
     2101 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagcgac
     2161 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2221 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2281 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2341 ggaagacaat agcaggcatg actagtccac tccctctctg cgcgctcgct cgctcactga
     2401 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     2461 gcgagcgcgc agagagggac agatccgggc ccgcatgcgt cgacaattca ctggccgtcg
     2521 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     2581 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     2641 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     2701 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     2761 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     2821 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     2881 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     2941 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     3001 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3061 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3121 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3181 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3241 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3301 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3361 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3421 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3481 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3541 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3601 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3661 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3721 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3781 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3841 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3901 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3961 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4021 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4081 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4141 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4201 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4261 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4321 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4381 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4441 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4501 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4561 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4621 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4681 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4741 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4801 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4861 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     4921 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     4981 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     5041 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     5101 aatttcacac aggaaacagc tatgaccatg attacgccaa gctctcgaga tctag
//
