LOCUS       sequence-48248-e        8967 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 8967)
  AUTHORS   Glover CP, Bienemann AS, Heywood DJ, Cosgrave AS, Uney JB
  TITLE     Adenoviral-mediated, high-level, cell-specific transgene expression:
            a SYN1-WPRE cassette mediates increased transgene expression with no
            loss of neuron specificity.
  JOURNAL   Mol Ther. 2002 May;5(5 Pt 1):509-16.
  PUBMED    11991741
REFERENCE   2  (bases 1 to 8967)
  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: "Mol Ther.";
            date: "2002-05"; volume: "5(5 Pt 1)"; pages: "509-16"
COMMENT     Sequence Label: pXCX.CMV.EGFP.WPRE
FEATURES             Location/Qualifiers
     source          1..8967
                     /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(1350..2063,2064..2066,2067..2069))
                     /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(1353..2063,2064..2066,2067..2069))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1395..1416)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1744..1763
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2003..2024
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2066..2075
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2116..2135)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2117..2135)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2180..2383)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2209..2229)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2384..2763
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             6519..6710
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6704..6726)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6812..6952
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6867..6886
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6967..6984)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7138..7726)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7218..7237)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7897..8688,8689..8757))
                     /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     8520..8539
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8758..8862)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8930..8948
                     /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 gcggccgctt tacttgtaca gctcgtccat gccgagagtg
     1381 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
     1441 ttgctcaggg cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
     1501 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg
     1561 tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg
     1621 ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     1681 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     1741 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
     1801 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac
     1861 tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     1921 gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
     1981 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     2041 gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtggatcccg ggcccgcggt
     2101 accaagcttg ggtctcccta tagtgagtcg tattaatttc gataagccag taagcagtgg
     2161 gttctctagt tagccagaga gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     2221 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
     2281 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
     2341 accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga
     2401 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     2461 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     2521 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     2581 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     2641 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     2701 atgggctatg aactaatgac cccgtaattg attactatta ataactagtc aataatcaat
     2761 gtcaacgcgt atatctggcc cgtacatcgc tagangatct ggaaggtgct gaggtacgat
     2821 gagacccgca ccaggtgcag accctgcgag tgtggcggta aacatattag gaaccagcct
     2881 gtgatgctgg atgtgaccga ggagctgagg cccgatcact tggtgctggc ctgcacccgc
     2941 gctgagtttg gctctagcga tgaagataca gattgaggta ctgaaatgtg tgggcgtggc
     3001 ttaagggtgg gaaagaatat ataaggtggg ggtcttatgt agttttgtat ctgttttgca
     3061 gcagccgccg ccgccatgag caccaactcg tttgatggaa gcattgtgag ctcatatttg
     3121 acaacgcgca tgcccccatg ggccggggtg cgtcagaatg tgatgggctc cagcattgat
     3181 ggtcgccccg tcctgcccgc aaactctact accttgacct acgagaccgt gtctggaacg
     3241 ccgttggaga ctgcagcctc cgccgccgct tcagccgctg cagccaccgc ccgcgggatt
     3301 gtgactgact ttgctttcct gagcccgctt gcaagcagtg cagcttcccg ttcatccgcc
     3361 cgcgatgaca agttgacggc tcttttggca caattggatt ctttgacccg ggaacttaat
     3421 gtcgtttctc agcagctgtt ggatctgcgc cagcaggttt ctgccctgaa ggcttcctcc
     3481 cctcccaatg cggtttaaaa cataaataaa aaaccagact ctgtttggat ttggatcaag
     3541 caagtgtctt gctgtcttta tttaggggtt ttgcgcgcgc ggtaggcccg ggaccagcgg
     3601 tctcggtcgt tgagggtcct gtgtattttt tccaggacgt ggtaaaggtg actctggatg
     3661 ttcagataca tgggcataag cccgtctctg gggtggaggt agcaccactg cagagcttca
     3721 tgctgcgggg tggtgttgta gatgatccag tcgtagcagg agcgctgggc gtggtgccta
     3781 aaaatgtctt tcagtagcaa gctgattgcc aggggcaggc ccttggtgta agtgtttaca
     3841 aagcggttaa gctgggatgg gtgcatacgt ggggatatga gatgcatctt ggactgtatt
     3901 tttaggttgg ctatgttccc agccatatcc ctccggggat tcatgttgtg cagaaccacc
     3961 agcacagtgt atccggtgca cttgggaaat ttgtcatgta gcttagaagg aaatgcgtgg
     4021 aagaacttgg agacgccctt gtgacctcca agattttcca tgcattcgtc cataatgatg
     4081 gcaatgggcc cacgggcggc ggcctgggcg aagatatttc tgggatcact aacgtcatag
     4141 ttgtgttcca ggatgagatc gtcataggcc atttttacaa agcgcgggcg gagggtgcca
     4201 gactgcggta taatggttcc atccggccca ggggcgtagt taccctcaca gatttgcatt
     4261 tcccacgctt tgagttcaga tggggggatc atgtctacct gcggggcgat gaagaaaacg
     4321 gtttccgggg taggggagat cagctgggaa gaaagcaggt tcctgagcag ctgcgactta
     4381 ccgcagccgg tgggcccgta aatcacacct attaccgggt gcaactggta gttaagagag
     4441 ctgcagctgc cgtcatccct gagcaggggg gccacttcgt taagcatgtc cctgactcgc
     4501 atgttttccc tgaccaaatc cgccagaagg cgctcgccgc ccagcgatag cagttcttgc
     4561 aaggaagcaa agtttttcaa cggtttgaga ccgtccgccg taggcatgct tttgagcgtt
     4621 tgaccaagca gttccaggcg gtcccacagc tcggtcacct gctctacggc atctcgatcc
     4681 agcatatctc ctcgtttcgc gggttggggc ggctttcgct gtacggcagt agtcggtgct
     4741 cgtccagacg ggccagggtc atgtctttcc acgggcgcag ggtcctcgtc agcgtagtct
     4801 gggtcacggt gaaggggtgc gctccgggct gcgcgctggc cagggtgcgc ttgaggctgg
     4861 tcctgctggt gctgaagcgc tgccggtctt cgccctgcgc gtcggccagg tagcatttga
     4921 ccatggtgtc atagtccagc ccctccgcgg cgtggccctt ggcgcgcagc ttgcccttgg
     4981 aggaggcgcc gcacgagggg cagtgcagac ttttgagggc gtagagcttg ggcgcgagaa
     5041 ataccgattc cggggagtag gcatccgcgc cgcaggcccc gcagacggtc tcgcattcca
     5101 cgagccaggt gagctctggc cgttcggggt caaaaaccag gtttccccca tgctttttga
     5161 tgcgtttctt acctctggtt tccatgagcc ggtgtccacg ctcggtgacg aaaaggctgt
     5221 ccgtgtcccc gtatacagac ttgagaggcc tgtcctcgac cgatgccctt gagagccttc
     5281 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     5341 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc
     5401 gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc
     5461 ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag
     5521 caggccatta tcgccggcat ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg
     5581 acgcgaggct ggatggcctt ccccattatg attcttctcg cttccggcgg catcgggatg
     5641 cccgcgttgc aggccatgct gtccaggcag gtagatgacg accatcaggg acagcttcaa
     5701 ggatcgctcg cggctcttac cagcctaact tcgatcactg gaccgctgat cgtcacggcg
     5761 atttatgccg cctcggcgag cacatggaac gggttggcat ggattgtagg cgccgcccta
     5821 taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga
     5881 atggaagccg gcggcacctc gctaacggat tcaccactcc aagaattgga gccaatcaat
     5941 tcttgcggag aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg
     6001 ccatctccag cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc
     6061 gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt
     6121 agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg
     6181 cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc
     6241 ggaagtcagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac
     6301 cctgtggaac acctacatct gtattaacga agcgctggca ttgaccctga gtgatttttc
     6361 tctggtcccg ccgcatccat accgccagtt gtttaccctc acaacgttcc agtaaccggg
     6421 catgttcatc atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta
     6481 cccccatgaa cagaaatccc ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg
     6541 cccttaacat ggcccgcttt atcagaagcc agacattaac gcttctggag aaactcaacg
     6601 agctggacgc ggatgaacag gcagacatct gtgaatcgct tcacgaccac gctgatgagc
     6661 tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     6721 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     6781 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     6841 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
     6901 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc
     6961 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7021 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7081 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7141 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7201 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7261 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7321 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7381 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7441 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7501 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7561 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7621 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7681 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7741 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7801 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7861 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7921 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7981 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8041 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8101 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8161 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat
     8221 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8281 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8341 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8401 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8461 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga
     8521 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8581 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8641 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8701 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8761 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8821 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8881 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     8941 cacgaggccc tttcgtcttc aagaatt
//
