LOCUS       sequence-466065-        5613 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 5613)
  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 5613)
  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: 
            HCT159_pAAV-eHGT_3Xcore_1137m-minBGprom-iCre297T-WPRE3-bGHpA
FEATURES             Location/Qualifiers
     source          1..5613
                     /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      1295..1304
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1307..1327
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2440..2460)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2647..2758
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2876..3016
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2876..3005
                     /label=AAV2 ITR
     rep_origin      3091..3546
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3178..3197)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3388..3409
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3563..3582)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3682..3704
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3742..3760)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3828..3932
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3933..4001,4002..4793)
                     /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(4151..4170)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4964..5552
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5453..5472
                     /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 gcgtgatttc tttgtctctg tggaagcact taagtgcagg
      181 ctttagttcc aatgacactc aggagcctct ggattccagc actggggatg ggggtggggt
      241 agaacgttct caggcctcac caacccctcc cctgtgtgct gcctttggga gagtcccaag
      301 gcttcagcat tacttaatta attaggcctc tactgctaca taggctcaga ttcaaaagaa
      361 cagagtggcc cacgtcagcc attcccggaa aagtctgatg gctggaagcc agaggactat
      421 gtgtctgcct tgctgccctt ggccagccca tcctgaatgc ccagactcgg acaatggagt
      481 aggtacagaa gggtagattt ctttgtctct gtggaagcac ttaagtgcag gctttagttc
      541 caatgacact caggagcctc tggattccag cactggggat gggggtgggg tagaacgttc
      601 tcaggcctca ccaacccctc ccctgtgtgc tgcctttggg agagtcccaa ggcttcagca
      661 ttacttaatt aattaggcct ctactgctac ataggctcag attcaaaaga acagagtggc
      721 ccacgtcagc cattcccgga aaagtctgat ggctggaagc cagaggacta tgtgtctgcc
      781 ttgctgccct tggccagccc atcctgaatg cccagactcg gacaatggag taggtacaga
      841 agggtagatt tctttgtctc tgtggaagca cttaagtgca ggctttagtt ccaatgacac
      901 tcaggagcct ctggattcca gcactgggga tgggggtggg gtagaacgtt ctcaggcctc
      961 accaacccct cccctgtgtg ctgcctttgg gagagtccca aggcttcagc attacttaat
     1021 taattaggcc tctactgcta cataggctca gattcaaaag aacagagtgg cccacgtcag
     1081 ccattcccgg aaaagtctga tggctggaag ccagaggact atgtgtctgc cttgctgccc
     1141 ttggccagcc catcctgaat gcccagactc ggacaatgga gtaggtacag aagggtacta
     1201 tttcactgag tttgagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta
     1261 catttgcttc tgggatccag atctttcgaa gctagccacc atggtgccca agaagaagag
     1321 gaaagtctcc aacctgctga ctgtgcacca aaacctgcct gccctccctg tggatgccac
     1381 ctctgatgaa gtcaggaaga acctgatgga catgttcagg gacaggcagg ccttctctga
     1441 acacacctgg aagatgctcc tgtctgtgtg cagatcctgg gctgcctggt gcaagctgaa
     1501 caacaggaaa tggttccctg ctgaacctga ggatgtgagg gactacctcc tgtacctgca
     1561 agccagaggc ctggctgtga agaccatcca acagcacctg ggccagctca acatgctgca
     1621 caggagatct ggcctgcctc gcccttctga ctccaatgct gtgtccctgg tgatgaggag
     1681 aatcagaaag gagaatgtgg atgctgggga gagagccaag caggccctgg cctttgaacg
     1741 cactgacttt gaccaagtca gatccctgat ggagaactct gacagatgcc aggacatcag
     1801 gaacctggcc ttcctgggca ttgcctacaa caccctgctg cgcattgccg aaattgccag
     1861 aatcagagtg aaggacatct cccgcaccga tggtgggaga atgctgatcc acattggcag
     1921 gaccaagacc ctggtgtcca cagctggtgt ggagaaggcc ctgtccctgg gggttaccaa
     1981 gctggtggag agatggatct ctgtgtctgg tgtggctgat gaccccaaca actacctgtt
     2041 ctgccgggtc agaaagaatg gtgtggctgc cccttctgcc acctcccaac tgtccacccg
     2101 ggccctggaa gggatctttg aggccaccca ccgcctgatc tatggtgcca aggatgactc
     2161 tgggcagaga tacctggcct ggtctggcca ctctgccaga gtgggtgctg ccaccgacat
     2221 ggccagggct ggtgtgtcca tccctgaaat catgcaggct ggtggctgga ccaatgtgaa
     2281 cattgtgatg aactacatca gaaacctgga ctctgagact ggggccatgg tgaggctgct
     2341 cgaagatggg gactgaggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga
     2401 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2461 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2521 ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct
     2581 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat
     2641 cttcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2701 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2761 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2821 aggattggga agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa
     2881 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     2941 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     3001 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3061 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     3121 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     3181 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3241 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3301 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3361 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3421 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     3481 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     3541 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     3601 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3661 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3721 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3781 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3841 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3901 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3961 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4021 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4081 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4141 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4201 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4261 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4321 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4381 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4441 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4501 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4561 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4621 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4681 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4741 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4801 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4861 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4921 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4981 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5041 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5101 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5161 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5221 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5281 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5341 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5401 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5461 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5521 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5581 tcctggcctt ttgctggcct tttgctcaca tgt
//
