LOCUS       sequence-48258-e        8823 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 8823)
  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 8823)
  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.TRE.EGFP.WPRE
FEATURES             Location/Qualifiers
     source          1..8823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..107
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     195..345
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
     polyA_signal    470..694
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     683..700
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    727..1315
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             833..844
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1242..1262
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1354..2067,2068..2070,2071..2073))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1357..2067,2068..2070,2071..2073))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1399..1420)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1748..1767
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2007..2028
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2070..2079
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2251..2275)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2272..2309)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    join(2342..2360,2361..2383,2384..2402,2403..2425,2426..2444,
                     2445..2467,2468..2486,2487..2509,2510..2528,2529..2551,
                     2552..2570,2571..2593,2594..2612)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     CDS             6379..6570
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6564..6586)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6672..6812
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6727..6746
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6827..6844)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6998..7586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7078..7097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7757..8548,8549..8617))
                     /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     8380..8399
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8618..8722)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8790..8808
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aattcatcat caataatata ccttattttg gattgaagcc aatatgataa tgagggggtg
       61 gagtttgtga cgtggcgcgg ggcgtgggaa cggggcgggt gacgtagtag tgtggcggaa
      121 gtgtgatgtt gcaagtgtgg cggaacacat gtaagcgacg gatgtggcaa aagtgacgtt
      181 tttggtgtgc gccggtgtac acaggaagtg acaattttcg cgcggtttta ggcggatgtt
      241 gtagtaaatt tgggcgtaac cgagtaagat ttggccattt tcgcgggaaa actgaataag
      301 aggaagtgaa atctgaataa ttttgtgtta ctcatagcgc gtaatatttg tctagggccg
      361 cggggacttt gaccgtttac gtggagactc gcccaggtgt ttttctcagg tgttttccgc
      421 gttccgggtc aaagttggcg ttttattatt atagtcagtc tagctggttc catagagccc
      481 accgcatccc cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
      541 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
      601 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
      661 caaacaacag atggctggca actagaaggc acagtcgagg ctgatcgtac cgagctcgaa
      721 ttccaggcgg ggaggcggcc caaagggaga tccgactcgt ctgagggcga aggcgaagac
      781 gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg aaggtccgct ggattgaggg
      841 ccgaagggac gtagcagaag gacgtcccgc gcagaatcca ggtggcaaca caggcgagca
      901 gccatggaaa ggacgtcagc ttccccgaca acaccacgga attgtcagtg cccaacagcc
      961 gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg gcaataggga
     1021 gggggaaagc gaaagtcccg gaaaggagct gacaggtggt ggcaatgccc caaccagtgg
     1081 gggttgcgtc agcaaacaca gtgcacacca cgccacgttg cctgacaacg ggccacaact
     1141 cctcataaag agacagcaac caggatttat acaaggagga gaaaatgaaa gccatacggg
     1201 aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta aaaggagcaa
     1261 catagttaag aataccagtc aatctttcac aaattttgta atccagaggt tgattgtcga
     1321 cggatcctct agatgcatgc tcgagcggcc gctttacttg tacagctcgt ccatgccgag
     1381 agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg
     1441 gtctttgctc agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc
     1501 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat
     1561 gttgtggcgg atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata
     1621 gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt
     1681 gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc
     1741 ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc
     1801 ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa
     1861 gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc
     1921 ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc
     1981 ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
     2041 cccggtgaac agctcctcgc ccttgctcac catggtggcg accggtggat cccgggcccg
     2101 cggtaccaag cttatcgatg cggccgcgct agcacgcgtc agctgactag aggatccccg
     2161 ggtaccgagc tcgaattcgg ggccgcggag gctggatcgg tcccggtgtc ttctatggag
     2221 gtcaaaacag cgtggatggc gtctccaggc gatctgacgg ttcactaaac gagctctgct
     2281 tatataggcc tcccaccgta cacgcctact cgacccgggt accgagctcg actttcactt
     2341 ttctctatca ctgataggga gtggtaaact cgactttcac ttttctctat cactgatagg
     2401 gagtggtaaa ctcgactttc acttttctct atcactgata gggagtggta aactcgactt
     2461 tcacttttct ctatcactga tagggagtgg taaactcgac tttcactttt ctctatcact
     2521 gatagggagt ggtaaactcg actttcactt ttctctatca ctgataggga gtggtaaact
     2581 cgactttcac ttttctctat cactgatagg gagtggtaaa ctcgacgcgt atatctggcc
     2641 cgtacatcgc tagangatct ggaaggtgct gaggtacgat gagacccgca ccaggtgcag
     2701 accctgcgag tgtggcggta aacatattag gaaccagcct gtgatgctgg atgtgaccga
     2761 ggagctgagg cccgatcact tggtgctggc ctgcacccgc gctgagtttg gctctagcga
     2821 tgaagataca gattgaggta ctgaaatgtg tgggcgtggc ttaagggtgg gaaagaatat
     2881 ataaggtggg ggtcttatgt agttttgtat ctgttttgca gcagccgccg ccgccatgag
     2941 caccaactcg tttgatggaa gcattgtgag ctcatatttg acaacgcgca tgcccccatg
     3001 ggccggggtg cgtcagaatg tgatgggctc cagcattgat ggtcgccccg tcctgcccgc
     3061 aaactctact accttgacct acgagaccgt gtctggaacg ccgttggaga ctgcagcctc
     3121 cgccgccgct tcagccgctg cagccaccgc ccgcgggatt gtgactgact ttgctttcct
     3181 gagcccgctt gcaagcagtg cagcttcccg ttcatccgcc cgcgatgaca agttgacggc
     3241 tcttttggca caattggatt ctttgacccg ggaacttaat gtcgtttctc agcagctgtt
     3301 ggatctgcgc cagcaggttt ctgccctgaa ggcttcctcc cctcccaatg cggtttaaaa
     3361 cataaataaa aaaccagact ctgtttggat ttggatcaag caagtgtctt gctgtcttta
     3421 tttaggggtt ttgcgcgcgc ggtaggcccg ggaccagcgg tctcggtcgt tgagggtcct
     3481 gtgtattttt tccaggacgt ggtaaaggtg actctggatg ttcagataca tgggcataag
     3541 cccgtctctg gggtggaggt agcaccactg cagagcttca tgctgcgggg tggtgttgta
     3601 gatgatccag tcgtagcagg agcgctgggc gtggtgccta aaaatgtctt tcagtagcaa
     3661 gctgattgcc aggggcaggc ccttggtgta agtgtttaca aagcggttaa gctgggatgg
     3721 gtgcatacgt ggggatatga gatgcatctt ggactgtatt tttaggttgg ctatgttccc
     3781 agccatatcc ctccggggat tcatgttgtg cagaaccacc agcacagtgt atccggtgca
     3841 cttgggaaat ttgtcatgta gcttagaagg aaatgcgtgg aagaacttgg agacgccctt
     3901 gtgacctcca agattttcca tgcattcgtc cataatgatg gcaatgggcc cacgggcggc
     3961 ggcctgggcg aagatatttc tgggatcact aacgtcatag ttgtgttcca ggatgagatc
     4021 gtcataggcc atttttacaa agcgcgggcg gagggtgcca gactgcggta taatggttcc
     4081 atccggccca ggggcgtagt taccctcaca gatttgcatt tcccacgctt tgagttcaga
     4141 tggggggatc atgtctacct gcggggcgat gaagaaaacg gtttccgggg taggggagat
     4201 cagctgggaa gaaagcaggt tcctgagcag ctgcgactta ccgcagccgg tgggcccgta
     4261 aatcacacct attaccgggt gcaactggta gttaagagag ctgcagctgc cgtcatccct
     4321 gagcaggggg gccacttcgt taagcatgtc cctgactcgc atgttttccc tgaccaaatc
     4381 cgccagaagg cgctcgccgc ccagcgatag cagttcttgc aaggaagcaa agtttttcaa
     4441 cggtttgaga ccgtccgccg taggcatgct tttgagcgtt tgaccaagca gttccaggcg
     4501 gtcccacagc tcggtcacct gctctacggc atctcgatcc agcatatctc ctcgtttcgc
     4561 gggttggggc ggctttcgct gtacggcagt agtcggtgct cgtccagacg ggccagggtc
     4621 atgtctttcc acgggcgcag ggtcctcgtc agcgtagtct gggtcacggt gaaggggtgc
     4681 gctccgggct gcgcgctggc cagggtgcgc ttgaggctgg tcctgctggt gctgaagcgc
     4741 tgccggtctt cgccctgcgc gtcggccagg tagcatttga ccatggtgtc atagtccagc
     4801 ccctccgcgg cgtggccctt ggcgcgcagc ttgcccttgg aggaggcgcc gcacgagggg
     4861 cagtgcagac ttttgagggc gtagagcttg ggcgcgagaa ataccgattc cggggagtag
     4921 gcatccgcgc cgcaggcccc gcagacggtc tcgcattcca cgagccaggt gagctctggc
     4981 cgttcggggt caaaaaccag gtttccccca tgctttttga tgcgtttctt acctctggtt
     5041 tccatgagcc ggtgtccacg ctcggtgacg aaaaggctgt ccgtgtcccc gtatacagac
     5101 ttgagaggcc tgtcctcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     5161 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     5221 cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag
     5281 cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc
     5341 cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat
     5401 ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt
     5461 ccccattatg attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct
     5521 gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac
     5581 cagcctaact tcgatcactg gaccgctgat cgtcacggcg atttatgccg cctcggcgag
     5641 cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct gcctccccgc
     5701 gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg gcggcacctc
     5761 gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag aactgtgaat
     5821 gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag cagccgcacg
     5881 cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt gctcctgtcg
     5941 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     6001 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     6061 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
     6121 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
     6181 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
     6241 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     6301 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
     6361 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     6421 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     6481 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
     6541 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     6601 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     6661 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     6721 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     6781 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     6841 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6901 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6961 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7021 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7081 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7141 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7201 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7261 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7321 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7381 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7441 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7501 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7561 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7621 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7681 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7741 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7801 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7861 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7921 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7981 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8041 ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt
     8101 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8161 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8221 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8281 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8341 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     8401 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8461 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8521 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8581 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8641 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8701 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     8761 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     8821 aag
//
