LOCUS       sequence-466752-        5773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing iCre297T AAV virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5773)
  AUTHORS   Kussick E, Johansen N, Taskin N, Chowdhury A, Quinlan MA, Fraser A, 
            Clark AG, Wynalda B, Martinez R, Groce EL, Reding M, Liang E, Shulga
            L, Huang C, Casper T, Clark M, Ho W, Gao Y, van Velthoven CTJ, 
            Sobieski C, Ferrer R, Berg MR, Curtis BC, English C, Day JC, Fortuna
            MG, Donadio N, Newman D, Yao S, Chakka AB, Goldy J, Torkelson A, 
            Guzman JB, Chakrabarty R, Nguy B, Guilford N, Pham TH, Wright V, 
            Ronellenfitch K, Naidoo R, Kenney J, Williford A, Ramakrishnan C, 
            Drinnenberg A, Gudsnuk K, Thyagarajan B, Smith KA, Dee N, Deisseroth
            K, Zeng H, Yao Z, Tasic B, Levi BP, Hodge R, Bakken TE, Lein ES, 
            Ting JT, Daigle TL
  TITLE     Enhancer AAVs for targeting spinal motor neurons and descending 
            motor pathways in rodents and macaque.
  JOURNAL   Cell Rep. 2025 Jun 24;44(6):115730. doi: 
            10.1016/j.celrep.2025.115730. Epub 2025 May 21.
  PUBMED    40403722
REFERENCE   2  (bases 1 to 5773)
  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.";
            date: "2025-06-24"; volume: "44(6):115730. doi"; pages: " 
            10.1016/j.celrep.2025.115730. Epub 2025 May 21"
COMMENT     Sequence Label: 
            HCT160_pAAV-3X-eHGT_1139m-minBGprom-iCre297T-WPRE3-bGHpA
FEATURES             Location/Qualifiers
     source          1..5773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1455..1464
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1467..1487
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2600..2620)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2807..2918
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3036..3176
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3036..3165
                     /label=AAV2 ITR
     rep_origin      3251..3706
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3338..3357)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3548..3569
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3723..3742)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3842..3864
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3902..3920)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3988..4092
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4093..4161,4162..4953)
                     /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(4311..4330)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5124..5712
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5613..5632
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaaatat tacaatacta aaaaaggaaa gacaagaaat
      181 gaagccagta gatatatttt caatagcata gtatatttcc agttaccaca tttttttgct
      241 gatgtttgta cttaaatgta tattaaaaag tttcttatcc caagtaaaga gcattactaa
      301 aattactgtg tgccactctg aaagatgaga gaaacccata ttcagtttcc ataatgaaat
      361 ccatatttgc ttggttgtga tccaaattgt ttttgaaaag tatatattta attgaggcag
      421 atgaagtggt ccaaaattta ataagtatta aaaaaaagga aaatttaaag agaaaatttc
      481 caattaaaga tgagtacctg atgattgctc aggcccaatt attaatattg attaatgttt
      541 tgtactttaa atattacaat actaaaaaag gaaagacaag aaatgaagcc agtagatata
      601 ttttcaatag catagtatat ttccagttac cacatttttt tgctgatgtt tgtacttaaa
      661 tgtatattaa aaagtttctt atcccaagta aagagcatta ctaaaattac tgtgtgccac
      721 tctgaaagat gagagaaacc catattcagt ttccataatg aaatccatat ttgcttggtt
      781 gtgatccaaa ttgtttttga aaagtatata tttaattgag gcagatgaag tggtccaaaa
      841 tttaataagt attaaaaaaa aggaaaattt aaagagaaaa tttccaatta aagatgagta
      901 cctgatgatt gctcaggccc aattattaat attgattaat gttttgtact ttaaatatta
      961 caatactaaa aaaggaaaga caagaaatga agccagtaga tatattttca atagcatagt
     1021 atatttccag ttaccacatt tttttgctga tgtttgtact taaatgtata ttaaaaagtt
     1081 tcttatccca agtaaagagc attactaaaa ttactgtgtg ccactctgaa agatgagaga
     1141 aacccatatt cagtttccat aatgaaatcc atatttgctt ggttgtgatc caaattgttt
     1201 ttgaaaagta tatatttaat tgaggcagat gaagtggtcc aaaatttaat aagtattaaa
     1261 aaaaaggaaa atttaaagag aaaatttcca attaaagatg agtacctgat gattgctcag
     1321 gcccaattat taatattgat taatgttttg tactttacta tttcactgag tttgagctcg
     1381 ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc tgggatccag
     1441 atctttcgaa gctagccacc atggtgccca agaagaagag gaaagtctcc aacctgctga
     1501 ctgtgcacca aaacctgcct gccctccctg tggatgccac ctctgatgaa gtcaggaaga
     1561 acctgatgga catgttcagg gacaggcagg ccttctctga acacacctgg aagatgctcc
     1621 tgtctgtgtg cagatcctgg gctgcctggt gcaagctgaa caacaggaaa tggttccctg
     1681 ctgaacctga ggatgtgagg gactacctcc tgtacctgca agccagaggc ctggctgtga
     1741 agaccatcca acagcacctg ggccagctca acatgctgca caggagatct ggcctgcctc
     1801 gcccttctga ctccaatgct gtgtccctgg tgatgaggag aatcagaaag gagaatgtgg
     1861 atgctgggga gagagccaag caggccctgg cctttgaacg cactgacttt gaccaagtca
     1921 gatccctgat ggagaactct gacagatgcc aggacatcag gaacctggcc ttcctgggca
     1981 ttgcctacaa caccctgctg cgcattgccg aaattgccag aatcagagtg aaggacatct
     2041 cccgcaccga tggtgggaga atgctgatcc acattggcag gaccaagacc ctggtgtcca
     2101 cagctggtgt ggagaaggcc ctgtccctgg gggttaccaa gctggtggag agatggatct
     2161 ctgtgtctgg tgtggctgat gaccccaaca actacctgtt ctgccgggtc agaaagaatg
     2221 gtgtggctgc cccttctgcc acctcccaac tgtccacccg ggccctggaa gggatctttg
     2281 aggccaccca ccgcctgatc tatggtgcca aggatgactc tgggcagaga tacctggcct
     2341 ggtctggcca ctctgccaga gtgggtgctg ccaccgacat ggccagggct ggtgtgtcca
     2401 tccctgaaat catgcaggct ggtggctgga ccaatgtgaa cattgtgatg aactacatca
     2461 gaaacctgga ctctgagact ggggccatgg tgaggctgct cgaagatggg gactgaggcg
     2521 cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat
     2581 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2641 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2701 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2761 ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt
     2821 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2881 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2941 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3001 aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     3061 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     3121 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     3181 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     3241 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3301 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     3361 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3421 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     3481 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3541 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     3601 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     3661 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     3721 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     3781 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     3841 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     3901 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     3961 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4021 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4081 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4141 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4201 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4261 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4321 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4381 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4441 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4501 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4561 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4621 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4681 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4741 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4801 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4861 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4921 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4981 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5041 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5101 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5161 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5221 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5281 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5341 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5401 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5461 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5521 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5581 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5641 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5701 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5761 tttgctcaca tgt
//
