LOCUS       sequence-48245-e        8804 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 8804)
  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 8804)
  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.Syn.EGFP.WPRE
FEATURES             Location/Qualifiers
     source          1..8804
                     /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"
     promoter        complement(2123..2570)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             6360..6551
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6545..6567)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6653..6793
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6708..6727
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6808..6825)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6979..7567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7059..7078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7738..8529,8530..8598))
                     /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     8361..8380
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8599..8703)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8771..8789
                     /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 ctctagagtc gactgcgctc tcaggcacga cacgactcct ccgctgccca
     2161 ccgcagactg aggcagcgct gagtcgccgg cgccgcagcg cagatggtcg cgcccgtgcc
     2221 cccctatctc gcgcctcgcg tggtgcggtc cggctgggcc ggcggcggcg cggacgcgac
     2281 caaggtggcc gggaagggga gtttgcgggg gaccggcgag tgacgtcagc gcgccttcag
     2341 tgctgaggcg gcggtggcgc gcgccgccag gcgggggcga aggcactgtc cgcggtgctg
     2401 aagctggcag tgcgcacgcg cctcgccgca tcctgtttcc cctccccctc tctgataggg
     2461 gatgcgcaat ttggggaatg ggggttgggt gcttgtccag tgggtcgggg tcggtcgtca
     2521 ggtaggcacc cccaccccgc ctcatcctgg tcctaaaacc cacttgcact catacgcagg
     2581 gccctctgca gctggcgcca tcgatacgcg tatatctggc ccgtacatcg ctagangatc
     2641 tggaaggtgc tgaggtacga tgagacccgc accaggtgca gaccctgcga gtgtggcggt
     2701 aaacatatta ggaaccagcc tgtgatgctg gatgtgaccg aggagctgag gcccgatcac
     2761 ttggtgctgg cctgcacccg cgctgagttt ggctctagcg atgaagatac agattgaggt
     2821 actgaaatgt gtgggcgtgg cttaagggtg ggaaagaata tataaggtgg gggtcttatg
     2881 tagttttgta tctgttttgc agcagccgcc gccgccatga gcaccaactc gtttgatgga
     2941 agcattgtga gctcatattt gacaacgcgc atgcccccat gggccggggt gcgtcagaat
     3001 gtgatgggct ccagcattga tggtcgcccc gtcctgcccg caaactctac taccttgacc
     3061 tacgagaccg tgtctggaac gccgttggag actgcagcct ccgccgccgc ttcagccgct
     3121 gcagccaccg cccgcgggat tgtgactgac tttgctttcc tgagcccgct tgcaagcagt
     3181 gcagcttccc gttcatccgc ccgcgatgac aagttgacgg ctcttttggc acaattggat
     3241 tctttgaccc gggaacttaa tgtcgtttct cagcagctgt tggatctgcg ccagcaggtt
     3301 tctgccctga aggcttcctc ccctcccaat gcggtttaaa acataaataa aaaaccagac
     3361 tctgtttgga tttggatcaa gcaagtgtct tgctgtcttt atttaggggt tttgcgcgcg
     3421 cggtaggccc gggaccagcg gtctcggtcg ttgagggtcc tgtgtatttt ttccaggacg
     3481 tggtaaaggt gactctggat gttcagatac atgggcataa gcccgtctct ggggtggagg
     3541 tagcaccact gcagagcttc atgctgcggg gtggtgttgt agatgatcca gtcgtagcag
     3601 gagcgctggg cgtggtgcct aaaaatgtct ttcagtagca agctgattgc caggggcagg
     3661 cccttggtgt aagtgtttac aaagcggtta agctgggatg ggtgcatacg tggggatatg
     3721 agatgcatct tggactgtat ttttaggttg gctatgttcc cagccatatc cctccgggga
     3781 ttcatgttgt gcagaaccac cagcacagtg tatccggtgc acttgggaaa tttgtcatgt
     3841 agcttagaag gaaatgcgtg gaagaacttg gagacgccct tgtgacctcc aagattttcc
     3901 atgcattcgt ccataatgat ggcaatgggc ccacgggcgg cggcctgggc gaagatattt
     3961 ctgggatcac taacgtcata gttgtgttcc aggatgagat cgtcataggc catttttaca
     4021 aagcgcgggc ggagggtgcc agactgcggt ataatggttc catccggccc aggggcgtag
     4081 ttaccctcac agatttgcat ttcccacgct ttgagttcag atggggggat catgtctacc
     4141 tgcggggcga tgaagaaaac ggtttccggg gtaggggaga tcagctggga agaaagcagg
     4201 ttcctgagca gctgcgactt accgcagccg gtgggcccgt aaatcacacc tattaccggg
     4261 tgcaactggt agttaagaga gctgcagctg ccgtcatccc tgagcagggg ggccacttcg
     4321 ttaagcatgt ccctgactcg catgttttcc ctgaccaaat ccgccagaag gcgctcgccg
     4381 cccagcgata gcagttcttg caaggaagca aagtttttca acggtttgag accgtccgcc
     4441 gtaggcatgc ttttgagcgt ttgaccaagc agttccaggc ggtcccacag ctcggtcacc
     4501 tgctctacgg catctcgatc cagcatatct cctcgtttcg cgggttgggg cggctttcgc
     4561 tgtacggcag tagtcggtgc tcgtccagac gggccagggt catgtctttc cacgggcgca
     4621 gggtcctcgt cagcgtagtc tgggtcacgg tgaaggggtg cgctccgggc tgcgcgctgg
     4681 ccagggtgcg cttgaggctg gtcctgctgg tgctgaagcg ctgccggtct tcgccctgcg
     4741 cgtcggccag gtagcatttg accatggtgt catagtccag cccctccgcg gcgtggccct
     4801 tggcgcgcag cttgcccttg gaggaggcgc cgcacgaggg gcagtgcaga cttttgaggg
     4861 cgtagagctt gggcgcgaga aataccgatt ccggggagta ggcatccgcg ccgcaggccc
     4921 cgcagacggt ctcgcattcc acgagccagg tgagctctgg ccgttcgggg tcaaaaacca
     4981 ggtttccccc atgctttttg atgcgtttct tacctctggt ttccatgagc cggtgtccac
     5041 gctcggtgac gaaaaggctg tccgtgtccc cgtatacaga cttgagaggc ctgtcctcga
     5101 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact
     5161 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     5221 gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg
     5281 tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc
     5341 accaaacgtt tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc
     5401 tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc
     5461 gcttccggcg gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac
     5521 gaccatcagg gacagcttca aggatcgctc gcggctctta ccagcctaac ttcgatcact
     5581 ggaccgctga tcgtcacggc gatttatgcc gcctcggcga gcacatggaa cgggttggca
     5641 tggattgtag gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg
     5701 agccgggcca cctcgacctg aatggaagcc ggcggcacct cgctaacgga ttcaccactc
     5761 caagaattgg agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc
     5821 agaacatatc catcgcgtcc gccatctcca gcagccgcac gcggcgcatc tcgggcagcg
     5881 ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct
     5941 ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg
     6001 actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg
     6061 tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca
     6121 tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc
     6181 attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct
     6241 cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct
     6301 ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag
     6361 tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa
     6421 cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc tgtgaatcgc
     6481 ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg
     6541 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     6601 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     6661 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
     6721 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6781 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6841 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6901 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6961 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7021 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7081 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7141 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7201 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7261 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7321 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7381 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7441 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7501 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7561 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7621 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7681 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7741 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7801 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7861 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7921 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7981 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8041 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8101 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8161 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8221 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8281 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8341 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8401 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8461 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8521 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8581 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8641 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8701 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     8761 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caag
//
