LOCUS       sequence-8168-ex        5365 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 5365)
  AUTHORS   Matsuda T, Cepko CL
  TITLE     Controlled expression of transgenes introduced by in vivo 
            electroporation.
  JOURNAL   Proc Natl Acad Sci U S A. 2007 Jan 16;104(3):1027-32. doi: 
            10.1073/pnas.0610155104
  PUBMED    17209010
REFERENCE   2  (bases 1 to 5365)
  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"; doi: "10.1073/pnas"; 
            journalName: "Proc Natl Acad Sci U S A"; date: "2007-01-16- 16"; 
            volume: "104"; issue: "3"; pages: "1027-32"
COMMENT     Sequence Label: pHes1-GFPd2
FEATURES             Location/Qualifiers
     source          1..5365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1176..1185
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1182..1184,1185..1187,1188..1898,1899..1904,
                     1905..2027)
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(1227..1248)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1488..1507)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1835..1856
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2153..2274
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2190..2209)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2244..2263
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2281..2736)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2418..2439)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2630..2649
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2763..2867
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2869..3226
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2869..3060
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2892..2912)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3077..3212
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3261..4055
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3261..4055
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3315..3334)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3925..3944
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4243..4262)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4287..4334
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4663..5251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5152..5171
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattac tagcgctacc ggactcagat ctacacgcac gcacacacac acatcctctg
       61 ggccccggcc gcagccgcgg cggcaataaa acatcctggc acgtgctccg actccaggcg
      121 cgccctcgcc ggccggctga tgtcaaactg cagctcggct ggtggagctc ttaaagggct
      181 cgcgcgcggg gcgccgaggc cgccttggcc gcccgcgggg tgaggagggg agagccccga
      241 ctcccttgcc cgccgtctat ccgtattgcc aactgggagc ctgcggggag aaaggtagag
      301 agagtggcgg aggaacgagc gcgcccctcc tgtcgcattg ctatcaaagc cccccttgtt
      361 catttttttt ttcgtttgaa attccatttc ctttctgccc agtaggaaat tattctgatt
      421 gttttctgga gaagcgtgag gtccaaggtt gcaattccaa gagaggaatg aatgggctag
      481 agggagacaa gcatccaaaa gacaccttgc tccgaaaacc ctggctgtga aggcatttgg
      541 ggggacggta agggcatgtt tagcgtgtgg ggtttagtat ctccccaacc tcatgagcag
      601 ttttgactcc cctaaacata cagagttcga gcgggaattc cggaagacat tagagtaaca
      661 catcctcttt accttgttcc ctcctttttt caatcactaa attttgtctt ggcctatatc
      721 tgttcaaaat atttttcaaa tgaacttatt atacaaagta gttatattgc atgcagcaag
      781 aacaataaaa accaaaggcc tggccacaaa agaaatagac tagactaaaa ctaagcaaag
      841 cccagaggaa agagttagca aagggttaaa atccttttga ttgacgttgt agcctccggt
      901 gccccgggct caggcgcgcg ccattggccg ccagaccttg tgcctagcgg ccaatggggg
      961 ggcgcagtcc acgagcggtg ccgcgtgtct cttcctccca ttggctgaaa gttactgtgg
     1021 gaaagaaagt ttgggaagtt tcacacgagc cgttcgcgtg cagtcccaga tatatataga
     1081 ggccgccagg gcctgcggat cacacaggat ctggagctgg tgctgataac agcggaatcc
     1141 cctgtctggt accgcgggcc cgggatccac cggtcgccac catggtgagc aagggcgagg
     1201 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1261 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1321 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1381 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1441 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1501 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1561 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1621 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     1681 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1741 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     1801 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1861 ccgccgggat cactctcggc atggacgagc tgtacaagaa gcttagccat ggcttcccgc
     1921 cggaggtgga ggagcaggat gatggcacgc tgcccatgtc ttgtgcccag gagagcggga
     1981 tggaccgtca ccctgcagcc tgtgcttctg ctaggatcaa tgtgtagatg cgcggccgcg
     2041 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2101 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2161 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2221 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     2281 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2341 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     2401 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2461 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     2521 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     2581 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     2641 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     2701 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     2761 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2821 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     2881 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2941 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3001 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3061 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3121 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3181 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3241 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3301 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3361 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3421 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     3481 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3541 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3601 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3661 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3721 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3781 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3841 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3901 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     3961 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4021 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4081 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4141 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4201 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4261 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     4321 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     4381 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     4441 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     4501 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4561 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4621 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4681 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4741 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4801 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4861 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4921 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4981 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5041 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5101 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5161 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5221 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5281 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5341 ggataaccgt attaccgcca tgcat
//
