LOCUS       sequence-461666-        6831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses diphtheria toxin A subunit in Cre-positive cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6831)
  AUTHORS   Tsutsui-Kimura I, Uchida N, Watabe-Uchida M
  TITLE     Dynamical management of potential threats regulated by dopamine and 
            direct- and indirect-pathway neurons in the tail of the striatum
  JOURNAL   bioRxiv 2022.02.05.479267
REFERENCE   2  (bases 1 to 6831)
  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: "bioRxiv 
            2022.02.05.479267"
COMMENT     Sequence Label: pAAV-CA-FLEX-dtA
FEATURES             Location/Qualifiers
     source          1..6831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        224..527
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          805..1821
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1745..1767
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1829..1848
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          2085..2150
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(2227..2799)
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     misc_feature    2958..3546
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3011..3031)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3429..3440)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3578..4054
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3777..3796)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4094..4234
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4094..4223
                     /label=AAV2 ITR
     rep_origin      4309..4764
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4396..4415)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4606..4627
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4781..4800)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4900..4922
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4960..4978)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5046..5150
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5151..5219,5220..6011)
                     /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(5369..5388)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6182..6770
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6671..6690
                     /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 gcgtaagctg ggctgcagga atctagttat taatagtaat
      181 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      241 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      301 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      361 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      421 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      481 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc
      541 ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat
      601 ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgccrggs
      661 ggggsggggs gggggggggg gggggggggg gsgrggsggr grggkgcggc ggcagccaat
      721 cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat
      781 aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc
      841 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
      901 agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt
      961 gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg
     1021 gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc
     1081 gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt
     1141 gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg
     1201 aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg
     1261 gtcgggctgc aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg
     1321 ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg
     1381 caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggrgggctc gggggagggg
     1441 cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt
     1501 ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg
     1561 aaatctggga ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc
     1621 ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc
     1681 cctctccagc ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg
     1741 gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat
     1801 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     1861 tttggcaaag aattgatttg atacgtacgg ttgaattcat tacctcgtat aggatacttt
     1921 atacgaagtt atgcagaatg gtagctggat tgtaagtact attagcaata tgaaacctct
     1981 taataacctc gtatagcata cattatacga agtaatagat ctaagcttga tctaggatct
     2041 gcattccacc actgctccca ttcatcagtt ccataggttg gaatctaaaa tacacaaaca
     2101 attagaatca gtagtttaac acattataca cttaaaaatt ttatatttac cttagagctt
     2161 taaatctctg taggtagttt gtccaattat gtcacaccac agaagtaagg ttccttcaca
     2221 aagagatcgc ctgacacgat ttcctgcaca ggcttgagcc atatactcat acatcgcatc
     2281 ttggccacgt tttccacggg tttcaaaatt aatctcaagt tctacgctta acgctttcgc
     2341 ctgttcccag ttattaatat attcaacgct agaactcccc tcagcgaagg gaaggctgag
     2401 cactacacgc gaagcaccat caccgaacct tttgataaac tcttccgttc cgacttgctc
     2461 catcaacggt tcagtgagac ttaaacctaa ctctttctta atagtttcgg cattatccac
     2521 ttttagtgcg agaaccttcg tcagtcctgg atacgtcact ttgaccacgc ctccagcttt
     2581 tccagagagc gggttttcat tatctacaga gtatcccgca gcgtcgtatt tattgtcggt
     2641 actataaaac cctttccaat catcgtcata atttccttgt gtaccagatt ttggcttttg
     2701 tatacctttt tgaatggaat ctacataacc aggtttagtc ccgtggtacg aagaaaagtt
     2761 ttccatcaca aaagatttag aagaatcaac aacatcatca ggatccatga tcgtcgacat
     2821 aacttcgtat aaagtatcct atacgaggta atttgcctta acccagaaat tatcagtact
     2881 attctttaga atggtgcaaa gaattacttc gtataatgta tgctatacga ggttatcgca
     2941 tgctagagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3001 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     3061 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     3121 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     3181 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     3241 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3301 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     3361 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     3421 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3481 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3541 ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc
     3601 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     3661 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     3721 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     3781 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     3841 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     3901 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     3961 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     4021 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
     4081 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4141 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4201 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     4261 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4321 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4381 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4441 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4501 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4561 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4621 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     4681 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4741 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4801 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4861 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4921 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4981 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     5041 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5101 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5161 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5221 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5281 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5341 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5401 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5461 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5521 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5581 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5641 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5701 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5761 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5821 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5881 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5941 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6001 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6061 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6121 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6181 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6241 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6301 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6361 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6421 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6481 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6541 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6601 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6661 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6721 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6781 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
