LOCUS       sequence-9422-ex        9537 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 9537)
  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 9537)
  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-1xGfaABC1D-EGFP
FEATURES             Location/Qualifiers
     source          1..9537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      718..727
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(724..726,727..729,730..1440)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(724..726,727..729,730..1440)
                     /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(769..790)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1030..1049)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1377..1398
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1441..1506
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(1633..1653)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2051..2062)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2365..2589
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2722..2741)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3762..3779)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3933..4521)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4013..4032)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4692..5483,5484..5552))
                     /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"
     promoter        complement(5553..5657)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5725..5743
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5794..5816)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5902..6040
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5955..5974
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7244..7264
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    8885..9118
                     /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 aacatatcct ggtgtggagt aggggacgct
       61 gctctgacag aggctcgggg gcctgagctg gctctgtgag ctggggagga ggcagacagc
      121 caggccttgt ctgcaagcag acctggcagc attgggctgg ccgcccccca gggcctcctc
      181 ttcatgccca gtgaatgact caccttggca cagacacaat gttcggggtg ggcacagtgc
      241 ctgcttcccg ccgcacccca gcccccctca aatgccttcc gagaagccca ttgagcaggg
      301 ggcttgcatt gcaccccagc ctgacagcct ggcatcttgg gataaaagca gcacagcccc
      361 ctaggggctg cccttgctgt gtggcgccac cggcggtgga gaacaaggct ctattcagcc
      421 tgtgcccagg aaaggggatc aggggatgcc caggcatgga cagtgggtgg caggggggga
      481 gaggagggct gtctgcttcc cagaagtcca aggacacaaa tgggtgaggg gagagctctc
      541 cccatagctg ggctgcggcc caaccccacc ccctcaggct atgccagggg gtgttgccag
      601 gggcacccgg gcatcgccag tctagcccac tccttcataa agccctcgca tcccaggagc
      661 gagcagagcc agagcaggtt ggagaggaga cgcatcacct ccgctgctcg cactagtgcc
      721 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      781 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      841 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      901 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
      961 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1021 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1081 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1141 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     1201 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     1261 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1321 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     1381 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1441 tccggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg
     1501 ggatccaccg gatctagata actgatcata atcagccata ccacgcggcc gctcccgtta
     1561 tttgcgctct gttcctgtta atcaacctct ggattacaaa atttgtgaaa gattgactgg
     1621 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     1681 tcatgctatt acttcccgta cggctttcat tttctcctcc ttgtataaat cctggttgct
     1741 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     1801 tgctgacgca acccccactg gttggggcat tgccaccacc tatcaactcc tttccgggac
     1861 tttcgctttc cccctcccta ttgccacggc ggaactcatt gccgcctgcc ttgcccgctg
     1921 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     1981 gtcctttcca tggctgctcg cctgtgttgc caactggatt ctgcgcggga cgtccttctg
     2041 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggttct
     2101 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2161 ctatcgatta gggataacag ggtaatggta cctttaagac caatgactta caaggcagct
     2221 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccag
     2281 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     2341 tgcttaagcc tcaataaagc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2401 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2461 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2521 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2581 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     2641 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2701 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2761 ccggctgtgc gggagaacgg agttctatta tgactcaaat cagtctcccc aagcattcgg
     2821 ggatcagagt ttttaaggat aacttagtgt gtagggggcc agtgagttgg agatgaaagc
     2881 gtagggagtc gaaggtgtcc ttttgcgccg agtcagttcc tgggtggggg ccacaagatc
     2941 ggatgagcca gtttatcaat ccgggggtgc cagctgatcc atggagtgca gggtctgcaa
     3001 aatatctcaa gcactgattg atcttaggtt ttacaatagt gatgttaccc caggaacaat
     3061 ttggggaagg tcagaatctt gtagcctgta gctgcatgac tcctaaacca taatttcttt
     3121 tttgtttttt tttttttatt tttgagacag ggtctcactc tgtcacctag gctggagtgc
     3181 agtggtgcaa tcacagctca ctgcagcctc aacgtcgtaa gctcaagcga tcctcccacc
     3241 tcagcctgcc tggtagctga gactacaagc gacgccccag ttaatttttg tatttttggt
     3301 agaggcagcg ttttgccgtg tggccctggc tggtctcgaa ctcctgggct caagtgatcc
     3361 agcctcagcc tcccaaagtg ctgggacaac cggggccagt cactgcacct ggccctaaac
     3421 cataatttct aatcttttgg ctaatttgtt agtcctacaa aggcagtcta gtccccaggc
     3481 aaaaaggggg tttgtttcgg gaaagggctg ttactgtctt tgtttcaaac tataaactaa
     3541 gttcctccta aacttagttc ggcctacacc caggaatgaa caaggagagc ttggaggtta
     3601 gaagcacgat ggaattggtt aggtcagatc tctttcactg tctgagttat aattttgcaa
     3661 tggtggttca aagactgccc gcttctgaca ccagtcgctg cattaatgaa tcggccaacg
     3721 cgcggggaga ggcggtttgc gtattggcgc tcttccgctt cctcgctcac tgactcgctg
     3781 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3841 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3901 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3961 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4021 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4081 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     4141 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4201 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4261 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4321 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     4381 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4441 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4501 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4561 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4621 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4681 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4741 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4801 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4861 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4921 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4981 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5041 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5101 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5161 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5221 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5281 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5341 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5401 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5461 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5521 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5581 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5641 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     5701 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa
     5761 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     5821 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     5881 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     5941 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     6001 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     6061 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agcgcgggga
     6121 ggcagagatt gcagtaagct gagatcgcag cactgcactc cagcctgggc gacagagtaa
     6181 gactctgtct caaaaataaa ataaataaat caatcagata ttccaatctt ttcctttatt
     6241 tatttattta ttttctattt tggaaacaca gtccttcctt attccagaat tacacatata
     6301 ttctattttt ctttatatgc tccagttttt tttagacctt cacctgaaat gtgtgtatac
     6361 aaaatctagg ccagtccagc agagcctaaa ggtaaaaaat aaaataataa aaaataaata
     6421 aaatctagct cactccttca catcaaaatg gagatacagc tgttagcatt aaataccaaa
     6481 taacccatct tgtcctcaat aattttaagc gcctctctcc accacatcta actcctgtca
     6541 aaggcatgtg ccccttccgg gcgctctgct gtgctgccaa ccaactggca tgtggactct
     6601 gcagggtccc taactgccaa gccccacagt gtgccctgag gctgcccctt ccttctagcg
     6661 gctgccccca ctcggctttg ctttccctag tttcagttac ttgcgttcag ccaaggtctg
     6721 aaactaggtg cgcacagagc ggtaagactg cgagagaaag agaccagctt tacagggggt
     6781 ttatcacagt gcaccctgac agtcgtcagc ctcacagggg gtttatcaca ttgcaccctg
     6841 acagtcgtca gcctcacagg gggtttatca cagtgcaccc ttacaatcat tccatttgat
     6901 tcacaatttt tttagtctct actgtgccta acttgtaagt taaatttgat cagaggtgtg
     6961 ttcccagagg ggaaaacagt atatacaggg ttcagtacta tcgcatttca gggtggcccg
     7021 agagctgcat ccggagtatc tagatggagt tccgcgttac ataacttacg gtaaatggcc
     7081 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgact gatgttccca
     7141 tagtaacgcc aatagggact ttccattgac gctaatggga gtttgttttg gcaccaaaat
     7201 caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg
     7261 cgtgtactct agaaggtcta tataagcaga gctcgtttag tgaaccgttt atactactta
     7321 tctggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     7381 cactgcttaa gcctcaataa agcttgcctt gagtgctcaa agtagtgtgt gcccgtctgt
     7441 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     7501 gcagtggcgc ccgaacaggg acttgaaagc gaaagtaaag ccagaggaga tctctcgacg
     7561 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     7621 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcggtat
     7681 taagcggggg agaattagat aaatgggaaa aaattcggtt aaggccaggg ggaaagaaac
     7741 aatataaact aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     7801 ctggcctttt agagacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     7861 ttcagacagg atcagaagaa cttagatcat tatataatac aatagcagtc ctctattgtg
     7921 tgcatcaaag gatagatgta aaagacacca aggaagcctt agataagata gaggaagagc
     7981 aaaacaaaag taagaaaaag gcacagcaag cagcagctga cacaggaaac aacagccagg
     8041 tcagccaaaa ttaccctata gtgcagaacc tccaggggca aatggtacat caggccatat
     8101 cacctagaac tttaaatgca tgggtaaaag tagtagaaga gaaggctttc agcccagaag
     8161 taatacccat gttttcagca ttatcagaag gagccacccc acaagattta aataccatgc
     8221 taaacacagt ggggggacat caagcagcca tgcaaatgtt aaaagagacc atcaatgagg
     8281 aagctgcaga atgggataga ttgcatccag tgcatgcagg gcctattgca ccaggccaga
     8341 tgagagaacc aaggggaagt gacatagcag gaactactag caaattaaga gaacaatttg
     8401 gaaataataa aacaataatc tttaagcaat cctcaggagg ggacccagaa attgtaacgc
     8461 acagttttaa ttgtggaggg gaatttttct actgtaattc aacacaactg tttaatagta
     8521 cttggtttaa tagtacttgg agtactgaag ggtcaaataa cactgaagga agtgacacaa
     8581 tcacactccc atgcagaata aaacaattta taaacatgtg gcaggaagta ggaaaagcaa
     8641 tgtatgcccc tcccatcagt ggacaaatta gatgttcatc aaatattact gggctgctat
     8701 taacaagaga tggtggtaat aacaacaatg ggtccgagat cttcagacct ggaggaggcg
     8761 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     8821 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     8881 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcacggt
     8941 caatgacgct gacggtacag gccagacaat tattgtctga tatagtgcag cagcagaaca
     9001 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     9061 aacagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     9121 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     9181 ggagtaataa atctctggaa cagatttgga ataacatgac ctggatggag tgggacagag
     9241 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     9301 aaaagaatga acaagaatta ttggaattag ataaatgggc ttgcggccgg aattcacaaa
     9361 tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg
     9421 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
     9481 ctaaaaattc naaaattttc gggtttatta cagggacagc agagatccac tttgggc
//
