LOCUS       sequence-325673-        5037 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 5037)
  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 5037)
  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..5037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    176..287
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     467..487
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1603..1623)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2358..2374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2358..2374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2371..2393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2382..2398
                     /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)."
     promoter        complement(join(2406..2412,2413..2430,2431..2436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2451..2472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2760..3348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2840..2859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3519..4310,4311..4379))
                     /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     4142..4161
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4380..4484)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4552..4570
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4608..4630)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4730..4749
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4943..4965
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4957..4974
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4958..4974
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
       61 agcgcgcaga gagggagtgg actagtcatg cctgctattg tcttcccaat cctccccctt
      121 gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc
      181 aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg
      241 cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg ctcgagccac
      301 ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc aggcggcgat
      361 gagttccgcc gtggcaagaa ctaaccagga tttatacaag gaggagaaaa tgaaagccat
      421 acgggaagca atagcatgat acaaaggcat taaagcagcg tatccacata gcgtaaaagg
      481 agcaacatag ttaagaatac cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt
      541 atgatatcga attcgcggcc gcggcgcgcc tcattagtcc ccatcctcga gcagcctcac
      601 catggcccca gtctcagagt ccaggtttct gatgtagttc atcacaatgt tcacattggt
      661 ccagccacca gcctgcatga tttcagggat ggacacacca gccctggcca tgtccctggc
      721 agcacccact ctggcagagt ggccagacca ggccaggtat ctctgcccag agtcatcctt
      781 ggcaccatag atcaggcggt gggtggcctc aaagatccct tccagggccc gggtggacag
      841 ttgggaggtg gcagaagggg cagccacacc attctttctg acccggcaga acaggtagtt
      901 gttggggtca tcagccacac cagacacaga gatccatctc tccaccagct tggtaacccc
      961 cagggacagg gccttctcca caccagctgt ggacaccagg gtcttggtcc tgccaatgtg
     1021 gatcagcatt ctcccaccat cggtgcggga gatgtccttc actctgattc tggcaatttc
     1081 ggcaatgcgc agcagggtgt tgtaggcaat gcccaggaag gccaggttcc tgatgtcctg
     1141 gcatctgtca gagttctcca tcagggatct gacttggtca aagtcagtgc gttcaaaggc
     1201 cagggcctgc ttggctctct ccccagcatc cacattctcc tttctgattc tcctcatcac
     1261 cagggacaca gcattggagt cagaagggcg aggcaggcca gatctcctgt gcagcatgtt
     1321 gagctggccc aggtgctgtt ggatggtctt cacagccagg cctctggctt gcaggtacag
     1381 gaggtagtcc ctcacatcct caggttcagc agggaaccat ttcctgttgt tcagcttgca
     1441 ccaggcagcc caggatctgc acacagacag gagcatcttc caggtgtgtt cagagaaggc
     1501 ctgcctgtcc ctgaacatgt ccatcaggtt cttcctgact tcatcagagg tggcatccac
     1561 agggagggca ggcaggtttt ggtgcacagt cagcaggttg gagactttcc tcttcttctt
     1621 gggcaccatg gtgctagctt cgaaggatcc cagaagcaaa tgtaagcaat agatggctct
     1681 gccctgactt ttatgcccag cccgagctcc ctactgtgaa actttgggtt cgtagctccc
     1741 caggatcaat tctgaacaaa gcctccagct gcagtgccat ccaatttgaa gcagacattg
     1801 gggacaattt aaggttttta tccacaagaa ggtttttttc cattctctta aatgcagcca
     1861 taattagagt aatttttcat gtagcccgct gattacagcg tttttaccgt caaagataat
     1921 tacctgtaat tttcttccac ttttaatact aaaaagccat ctttatttag attcaggaac
     1981 aggaaaggcg aaacaaaaga gggaaattat tctgttattc atacacaaat tgcagagacg
     2041 taggacctaa aattgaaaat taaccaaaat tataatgctg aaaagaatgg aagaggctgc
     2101 agaagtacag ctggtagtgg ggctttgctg aattattcaa attacgttag ggagaatgct
     2161 accatacatc gaaaccaaca ctaatttttc gttaaaaaaa attcaccaga tgcatgccaa
     2221 accactgtga gtgcatagag ctcggtacct atacgcgtcc taggtcacga ctcmrysmsy
     2281 smrsgarcsc syagwgaggg agtggcccag atccccccgg gaagctttct agatctcgag
     2341 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2401 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2461 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2521 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2581 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2641 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2701 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2761 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2821 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2881 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2941 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3001 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3061 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3121 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3181 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3241 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3301 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3361 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3421 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3481 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3541 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3601 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3661 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3721 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3781 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3841 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3901 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3961 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4021 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4081 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4141 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4201 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4261 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4321 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4381 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4441 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4501 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     4561 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4621 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4681 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     4741 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     4801 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     4861 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     4921 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg
     4981 tcgacgcatg cgggcccgga tctgtccctc tctgcgcgct cgctcgctca ctgaggc
//
