LOCUS       sequence-9421-ex        9383 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 9383)
  AUTHORS   Liu B, Paton JF, Kasparov S
  TITLE     Viral vectors based on bidirectional cell-specific mammalian 
            promoters and transcriptional amplification strategy for use in 
            vitro and in vivo.
  JOURNAL   BMC Biotechnol. 2008 May 16;8:49. doi: 10.1186/1472-6750-8-49.
  PUBMED    18485188
REFERENCE   2  (bases 1 to 9383)
  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: "BMC 
            Biotechnol."; date: "2008-05-16"; volume: "8:49. doi"; pages: " 
            10.1186/1472-6750-8-49"
COMMENT     Sequence Label: pTYF-1xSYN-EGFP
FEATURES             Location/Qualifiers
     source          1..9383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        66..513
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      564..573
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(570..572,573..575,576..1286)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(570..572,573..575,576..1286)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(615..636)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(876..895)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1223..1244
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1287..1352
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(1479..1499)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1897..1908)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2211..2435
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2568..2587)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3608..3625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3779..4367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3859..3878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4538..5329,5330..5398))
                     /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     5161..5180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5399..5503)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5571..5589
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5640..5662)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5748..5886
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5801..5820
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7090..7110
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    8731..8964
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 ggcctaggga taacagggta atgatcgcgt atcgatggcg ccagctgcag agggccctgc
       61 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      121 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      181 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      241 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      301 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      361 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      421 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      481 agcggaggag tcgtgtcgtg cctgagagcg cagtcgactc tagagctgca ggcaagcttg
      541 gtaccgagct cggatccact agtgccacca tggtgagcaa gggcgaggag ctgttcaccg
      601 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      661 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
      721 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
      781 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      841 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
      901 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
      961 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1021 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1081 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1141 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     1201 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1261 ctctcggcat ggacgagctg tacaagtccg gactcagatc tcgagctcaa gcttcgaatt
     1321 ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
     1381 gccataccac gcggccgctc ccgttatttg cgctctgttc ctgttaatca acctctggat
     1441 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     1501 ggatacgctg ctttaatgcc tttgtatcat gctattactt cccgtacggc tttcattttc
     1561 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     1621 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     1681 accacctatc aactcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     1741 ctcattgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     1801 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccaac
     1861 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     1921 ccttcccgcg gcctgctgcc ggttctgcgg cctcttccgc gtcttcgcct tcgccctcag
     1981 acgagtcgga tctccctttg ggccgcctat cgattaggga taacagggta atggtacctt
     2041 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     2101 gactggaagg gctaattcac tcccagggtc tctctggtta gaccagatct gagcctggga
     2161 gctctctggc taactaggga acccactgct taagcctcaa taaagctcga ctgtgccttc
     2221 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2281 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2341 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2401 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2461 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2521 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2581 cttcccttcc tttctcgcca cgttcgccgg ctgtgcggga gaacggagtt ctattatgac
     2641 tcaaatcagt ctccccaagc attcggggat cagagttttt aaggataact tagtgtgtag
     2701 ggggccagtg agttggagat gaaagcgtag ggagtcgaag gtgtcctttt gcgccgagtc
     2761 agttcctggg tgggggccac aagatcggat gagccagttt atcaatccgg gggtgccagc
     2821 tgatccatgg agtgcagggt ctgcaaaata tctcaagcac tgattgatct taggttttac
     2881 aatagtgatg ttaccccagg aacaatttgg ggaaggtcag aatcttgtag cctgtagctg
     2941 catgactcct aaaccataat ttcttttttg tttttttttt tttatttttg agacagggtc
     3001 tcactctgtc acctaggctg gagtgcagtg gtgcaatcac agctcactgc agcctcaacg
     3061 tcgtaagctc aagcgatcct cccacctcag cctgcctggt agctgagact acaagcgacg
     3121 ccccagttaa tttttgtatt tttggtagag gcagcgtttt gccgtgtggc cctggctggt
     3181 ctcgaactcc tgggctcaag tgatccagcc tcagcctccc aaagtgctgg gacaaccggg
     3241 gccagtcact gcacctggcc ctaaaccata atttctaatc ttttggctaa tttgttagtc
     3301 ctacaaaggc agtctagtcc ccaggcaaaa agggggtttg tttcgggaaa gggctgttac
     3361 tgtctttgtt tcaaactata aactaagttc ctcctaaact tagttcggcc tacacccagg
     3421 aatgaacaag gagagcttgg aggttagaag cacgatggaa ttggttaggt cagatctctt
     3481 tcactgtctg agttataatt ttgcaatggt ggttcaaaga ctgcccgctt ctgacaccag
     3541 tcgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tggcgctctt
     3601 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3661 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3721 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3781 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3841 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3901 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3961 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4021 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4081 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4141 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4201 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4261 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4321 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4381 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4441 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4501 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4561 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4621 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4681 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4741 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4801 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4861 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4921 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4981 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5041 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5101 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5161 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5221 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5281 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5341 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5401 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5461 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5521 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5581 tcacgaggcc ctttcgtctt caagaactgc ctcgcgcgtt tcggtgatga cggtgaaaac
     5641 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     5701 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     5761 cagtcacgta gcgatagcgg agtgtactgg cttaactatg cggcatcaga gcagattgta
     5821 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     5881 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     5941 tcttcgctat tacgccagcg cggggaggca gagattgcag taagctgaga tcgcagcact
     6001 gcactccagc ctgggcgaca gagtaagact ctgtctcaaa aataaaataa ataaatcaat
     6061 cagatattcc aatcttttcc tttatttatt tatttatttt ctattttgga aacacagtcc
     6121 ttccttattc cagaattaca catatattct atttttcttt atatgctcca gtttttttta
     6181 gaccttcacc tgaaatgtgt gtatacaaaa tctaggccag tccagcagag cctaaaggta
     6241 aaaaataaaa taataaaaaa taaataaaat ctagctcact ccttcacatc aaaatggaga
     6301 tacagctgtt agcattaaat accaaataac ccatcttgtc ctcaataatt ttaagcgcct
     6361 ctctccacca catctaactc ctgtcaaagg catgtgcccc ttccgggcgc tctgctgtgc
     6421 tgccaaccaa ctggcatgtg gactctgcag ggtccctaac tgccaagccc cacagtgtgc
     6481 cctgaggctg ccccttcctt ctagcggctg cccccactcg gctttgcttt ccctagtttc
     6541 agttacttgc gttcagccaa ggtctgaaac taggtgcgca cagagcggta agactgcgag
     6601 agaaagagac cagctttaca gggggtttat cacagtgcac cctgacagtc gtcagcctca
     6661 cagggggttt atcacattgc accctgacag tcgtcagcct cacagggggt ttatcacagt
     6721 gcacccttac aatcattcca tttgattcac aattttttta gtctctactg tgcctaactt
     6781 gtaagttaaa tttgatcaga ggtgtgttcc cagaggggaa aacagtatat acagggttca
     6841 gtactatcgc atttcagggt ggcccgagag ctgcatccgg agtatctaga tggagttccg
     6901 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     6961 gacgtcaata atgactgatg ttcccatagt aacgccaata gggactttcc attgacgcta
     7021 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg
     7081 ccccgttgac gcaaatgggc ggtaggcgtg tactctagaa ggtctatata agcagagctc
     7141 gtttagtgaa ccgtttatac tacttatctg gtctctctgg ttagaccaga tctgagcctg
     7201 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     7261 gctcaaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     7321 cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa
     7381 gtaaagccag aggagatctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga
     7441 ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag
     7501 agagatgggt gcgagagcgt cggtattaag cgggggagaa ttagataaat gggaaaaaat
     7561 tcggttaagg ccagggggaa agaaacaata taaactaaaa catatagtat gggcaagcag
     7621 ggagctagaa cgattcgcag ttaatcctgg ccttttagag acatcagaag gctgtagaca
     7681 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     7741 taatacaata gcagtcctct attgtgtgca tcaaaggata gatgtaaaag acaccaagga
     7801 agccttagat aagatagagg aagagcaaaa caaaagtaag aaaaaggcac agcaagcagc
     7861 agctgacaca ggaaacaaca gccaggtcag ccaaaattac cctatagtgc agaacctcca
     7921 ggggcaaatg gtacatcagg ccatatcacc tagaacttta aatgcatggg taaaagtagt
     7981 agaagagaag gctttcagcc cagaagtaat acccatgttt tcagcattat cagaaggagc
     8041 caccccacaa gatttaaata ccatgctaaa cacagtgggg ggacatcaag cagccatgca
     8101 aatgttaaaa gagaccatca atgaggaagc tgcagaatgg gatagattgc atccagtgca
     8161 tgcagggcct attgcaccag gccagatgag agaaccaagg ggaagtgaca tagcaggaac
     8221 tactagcaaa ttaagagaac aatttggaaa taataaaaca ataatcttta agcaatcctc
     8281 aggaggggac ccagaaattg taacgcacag ttttaattgt ggaggggaat ttttctactg
     8341 taattcaaca caactgttta atagtacttg gtttaatagt acttggagta ctgaagggtc
     8401 aaataacact gaaggaagtg acacaatcac actcccatgc agaataaaac aatttataaa
     8461 catgtggcag gaagtaggaa aagcaatgta tgcccctccc atcagtggac aaattagatg
     8521 ttcatcaaat attactgggc tgctattaac aagagatggt ggtaataaca acaatgggtc
     8581 cgagatcttc agacctggag gaggcgatat gagggacaat tggagaagtg aattatataa
     8641 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     8701 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     8761 agcaggaagc actatgggcg cacggtcaat gacgctgacg gtacaggcca gacaattatt
     8821 gtctgatata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     8881 gttgcaactc acagtctggg gcatcaaaca gctccaggca agaatcctgg ctgtggaaag
     8941 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     9001 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaataa
     9061 catgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
     9121 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
     9181 atgggcttgc ggccggaatt cacaaatggc agtattcatc cacaatttta aaagaaaagg
     9241 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     9301 aactaaagaa ttacaaaaac aaattactaa aaattcnaaa attttcgggt ttattacagg
     9361 gacagcagag atccactttg ggc
//
