LOCUS       sequence-48257-e        9415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9415)
  AUTHORS   Lee YB, Cosgrave AS, Glover CP, Bienemann A, Heywood D, Hobson RJ, 
            Uney JB
  TITLE     Increased utility in the CNS of a powerful neuron-specific 
            tetracycline-regulatable adenoviral system developed using a 
            post-transcriptional enhancer.
  JOURNAL   J Gene Med. 2005 May;7(5):576-83.
  PUBMED    15580589
REFERENCE   2  (bases 1 to 9415)
  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: "J Gene 
            Med."; date: "2005-05"; volume: "7(5)"; pages: "576-83"
COMMENT     Sequence Label: pXCX.CMV.tTA.WPRE
FEATURES             Location/Qualifiers
     source          1..9415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..103
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     191..341
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
     polyA_signal    466..690
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     679..696
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    723..1311
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             829..840
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1238..1258
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1414..1800,1801..2421))
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     2329..2348
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(2444..2460)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2445..2461)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2550..2569)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2551..2569)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        2726..3105
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             6967..7158
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(7152..7174)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7260..7400
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7315..7334
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7415..7432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7586..8174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7666..7685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8345..9136,9137..9205))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8968..8987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9206..9310)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9378..9396
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 catcatcaat aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt
       61 ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt
      121 gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg
      181 gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag
      241 taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga
      301 agtgaaatct gaataatttt gtgttactca tagcgcgtaa tatttgtcta gggccgcggg
      361 gactttgacc gtttacgtgg agactcgccc aggtgttttt ctcaggtgtt ttccgcgttc
      421 cgggtcaaag ttggcgtttt attattatag tcagtctagc tggttccata gagcccaccg
      481 catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc
      541 caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt
      601 aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa
      661 caacagatgg ctggcaacta gaaggcacag tcgaggctga tcgtaccgag ctcgaattcc
      721 aggcggggag gcggcccaaa gggagatccg actcgtctga gggcgaaggc gaagacgcgg
      781 aagaggccgc agagccggca gcaggccgcg ggaaggaagg tccgctggat tgagggccga
      841 agggacgtag cagaaggacg tcccgcgcag aatccaggtg gcaacacagg cgagcagcca
      901 tggaaaggac gtcagcttcc ccgacaacac cacggaattg tcagtgccca acagccgagc
      961 ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa tagggagggg
     1021 gaaagcgaaa gtcccggaaa ggagctgaca ggtggtggca atgccccaac cagtgggggt
     1081 tgcgtcagca aacacagtgc acaccacgcc acgttgcctg acaacgggcc acaactcctc
     1141 ataaagagac agcaaccagg atttatacaa ggaggagaaa atgaaagcca tacgggaagc
     1201 aatagcatga tacaaaggca ttaaagcagc gtatccacat agcgtaaaag gagcaacata
     1261 gttaagaata ccagtcaatc tttcacaaat tttgtaatcc agaggttgat tgtcgacgga
     1321 tcctctagat gcatgctcga gcggccgcca gtgtgatgga atctgcagaa ttccagcaca
     1381 ctggcggccg ttactagtgg atcctcgcgc cccctaccca ccgtactcgt caattccaag
     1441 ggcatcggta aacatctgct caaactcgaa gtcggccata tccagagcgc cgtagggggc
     1501 ggagtcgtgg ggggtaaatc ccggacccgg ggaatccccg tcccccaaca tgtccagatc
     1561 gaaatcgtct agcgcgtcgg catgcgccat cgccacgtcc tcgccgtcta agtggagctc
     1621 gtcccccagg ctgacatcgg tcgggggggc cgtcgacagt ctgcgcgtgt gtcccgcggg
     1681 gagaaaggac aggcgcggag ccgccagccc cgcctcttcg ggggcgtcgt cgtccgggag
     1741 atcgagcagg ccctcgatgg tagacccgta attgtttttc gtacgcgcgc ggctgtacgc
     1801 ggacccactt tcacatttaa gttgtttttc taatccgcat atgatcaatt caaggccgaa
     1861 taagaaggct ggctctgcac cttggtgatc aaataattcg atagcttgtc gtaataatgg
     1921 cggcatacta tcagtagtag gtgtttccct ttcttcttta gcgacttgat gctcttgatc
     1981 ttccaatacg caacctaaag taaaatgccc cacagcgctg agtgcatata atgcattctc
     2041 tagtgaaaaa ccttgttggc ataaaaaggc taattgattt tcgagagttt catactgttt
     2101 ttctgtaggc cgtgtaccta aatgtacttt tgctccatcg cgatgactta gtaaagcaca
     2161 tctaaaactt ttagcgttat tacgtaaaaa atcttgccag ctttcccctt ctaaagggca
     2221 aaagtgagta tggtgcctat ctaacatctc aatggctaag gcgtcgagca aagcccgctt
     2281 attttttaca tgccaataca atgtaggctg ctctacacct agcttctggg cgagtttacg
     2341 ggttgttaaa ccttcgattc cgacctcatt aagcagctct aatgcgctgt taatcacttt
     2401 acttttatct aatctagaca tatgaattcg atatcaagct tatcgatacc gtcgacctcg
     2461 agcggccgcc agtgtgatgg aatctgcaga attccagcac actggcggcc gttactagtg
     2521 gatccgagct cggtaccaag cttgggtctc cctatagtga gtcgtattaa tttcgataag
     2581 ccagtaagca gtgggttctc tagtagtccc gttgattttg gtgccaaaac aaactcccat
     2641 tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat
     2701 tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact
     2761 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     2821 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     2881 gggcagttta gcgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     2941 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
     3001 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
     3061 accccgtaat tgattactat taataactag tcaataatca atgtcaacat ggcggtaatg
     3121 ttggacatga gccaatataa atgtacatat tatgatatgg atacaacgta tgcaatgggc
     3181 caagctcctc gagcggccgc cagtgtgatg gatcgcgtat atctggcccg tacatcgcta
     3241 gangatctgg aaggtgctga ggtacgatga gacccgcacc aggtgcagac cctgcgagtg
     3301 tggcggtaaa catattagga accagcctgt gatgctggat gtgaccgagg agctgaggcc
     3361 cgatcacttg gtgctggcct gcacccgcgc tgagtttggc tctagcgatg aagatacaga
     3421 ttgaggtact gaaatgtgtg ggcgtggctt aagggtggga aagaatatat aaggtggggg
     3481 tcttatgtag ttttgtatct gttttgcagc agccgccgcc gccatgagca ccaactcgtt
     3541 tgatggaagc attgtgagct catatttgac aacgcgcatg cccccatggg ccggggtgcg
     3601 tcagaatgtg atgggctcca gcattgatgg tcgccccgtc ctgcccgcaa actctactac
     3661 cttgacctac gagaccgtgt ctggaacgcc gttggagact gcagcctccg ccgccgcttc
     3721 agccgctgca gccaccgccc gcgggattgt gactgacttt gctttcctga gcccgcttgc
     3781 aagcagtgca gcttcccgtt catccgcccg cgatgacaag ttgacggctc ttttggcaca
     3841 attggattct ttgacccggg aacttaatgt cgtttctcag cagctgttgg atctgcgcca
     3901 gcaggtttct gccctgaagg cttcctcccc tcccaatgcg gtttaaaaca taaataaaaa
     3961 accagactct gtttggattt ggatcaagca agtgtcttgc tgtctttatt taggggtttt
     4021 gcgcgcgcgg taggcccggg accagcggtc tcggtcgttg agggtcctgt gtattttttc
     4081 caggacgtgg taaaggtgac tctggatgtt cagatacatg ggcataagcc cgtctctggg
     4141 gtggaggtag caccactgca gagcttcatg ctgcggggtg gtgttgtaga tgatccagtc
     4201 gtagcaggag cgctgggcgt ggtgcctaaa aatgtctttc agtagcaagc tgattgccag
     4261 gggcaggccc ttggtgtaag tgtttacaaa gcggttaagc tgggatgggt gcatacgtgg
     4321 ggatatgaga tgcatcttgg actgtatttt taggttggct atgttcccag ccatatccct
     4381 ccggggattc atgttgtgca gaaccaccag cacagtgtat ccggtgcact tgggaaattt
     4441 gtcatgtagc ttagaaggaa atgcgtggaa gaacttggag acgcccttgt gacctccaag
     4501 attttccatg cattcgtcca taatgatggc aatgggccca cgggcggcgg cctgggcgaa
     4561 gatatttctg ggatcactaa cgtcatagtt gtgttccagg atgagatcgt cataggccat
     4621 ttttacaaag cgcgggcgga gggtgccaga ctgcggtata atggttccat ccggcccagg
     4681 ggcgtagtta ccctcacaga tttgcatttc ccacgctttg agttcagatg gggggatcat
     4741 gtctacctgc ggggcgatga agaaaacggt ttccggggta ggggagatca gctgggaaga
     4801 aagcaggttc ctgagcagct gcgacttacc gcagccggtg ggcccgtaaa tcacacctat
     4861 taccgggtgc aactggtagt taagagagct gcagctgccg tcatccctga gcaggggggc
     4921 cacttcgtta agcatgtccc tgactcgcat gttttccctg accaaatccg ccagaaggcg
     4981 ctcgccgccc agcgatagca gttcttgcaa ggaagcaaag tttttcaacg gtttgagacc
     5041 gtccgccgta ggcatgcttt tgagcgtttg accaagcagt tccaggcggt cccacagctc
     5101 ggtcacctgc tctacggcat ctcgatccag catatctcct cgtttcgcgg gttggggcgg
     5161 ctttcgctgt acggcagtag tcggtgctcg tccagacggg ccagggtcat gtctttccac
     5221 gggcgcaggg tcctcgtcag cgtagtctgg gtcacggtga aggggtgcgc tccgggctgc
     5281 gcgctggcca gggtgcgctt gaggctggtc ctgctggtgc tgaagcgctg ccggtcttcg
     5341 ccctgcgcgt cggccaggta gcatttgacc atggtgtcat agtccagccc ctccgcggcg
     5401 tggcccttgg cgcgcagctt gcccttggag gaggcgccgc acgaggggca gtgcagactt
     5461 ttgagggcgt agagcttggg cgcgagaaat accgattccg gggagtaggc atccgcgccg
     5521 caggccccgc agacggtctc gcattccacg agccaggtga gctctggccg ttcggggtca
     5581 aaaaccaggt ttcccccatg ctttttgatg cgtttcttac ctctggtttc catgagccgg
     5641 tgtccacgct cggtgacgaa aaggctgtcc gtgtccccgt atacagactt gagaggcctg
     5701 tcctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     5761 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     5821 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
     5881 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
     5941 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc
     6001 gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat
     6061 tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt
     6121 agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc
     6181 gatcactgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg
     6241 gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg
     6301 tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc
     6361 accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac
     6421 ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg
     6481 ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg
     6541 ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg
     6601 tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt
     6661 ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg
     6721 atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag
     6781 cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccagttgt
     6841 ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg tatcgtgagc
     6901 atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc ttacacggag
     6961 gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag
     7021 acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt
     7081 gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat
     7141 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     7201 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     7261 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
     7321 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     7381 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     7441 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     7501 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     7561 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     7621 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     7681 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     7741 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     7801 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     7861 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     7921 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     7981 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     8041 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     8101 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     8161 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     8221 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     8281 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     8341 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     8401 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     8461 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     8521 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     8581 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     8641 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     8701 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8761 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8821 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     8881 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8941 gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag
     9001 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     9061 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     9121 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     9181 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     9241 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     9301 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     9361 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaatt
//
