LOCUS       sequence-181057-        8068 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 8068)
  AUTHORS   Gerrard L, Zhao D, Clark AJ, Cui W
  TITLE     Stably transfected human embryonic stem cell clones express 
            OCT4-specific green fluorescent protein and maintain self-renewal 
            and pluripotency.
  JOURNAL   Stem Cells. 2005;23(1):124-33.
  PUBMED    15625129
REFERENCE   2  (bases 1 to 8068)
  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: "Stem 
            Cells."; date: "2005"; volume: "23(1)"; pages: "124-33"
COMMENT     Sequence Label: phOct4-EGFP
FEATURES             Location/Qualifiers
     source          1..8068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      4062..4071
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4068..4070,4071..4073,4074..4787)
                     /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             join(4068..4070,4071..4073,4074..4784)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4113..4134)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4374..4393)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4721..4742
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4910..5031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4947..4966)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5001..5020
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5038..5493)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5175..5196)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5387..5406
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5520..5624
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5626..5983
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5626..5817
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5649..5669)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5834..5969
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6018..6812
                     /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             6018..6812
                     /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(6072..6091)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6682..6701
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7000..7019)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7044..7091
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7420..8008
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7909..7928
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
       61 ttactagcgc taccggactc agatctcgag ctcaagcttc ccatgtcaag taagtggggt
      121 gggtagatgg gagggcatat gagagcgggg cagcaatttt atgtcttagt tggaaggaag
      181 tttagagttc attgagtcca aatcctcttt actaggtgag gaaactgagg cacaggaggg
      241 ggaggtgcca ttagccaggg agtcagagcc aggacttaga acctggtcca tgcctcctag
      301 tttgttcctg accaatcctc ccagctcacc cactcctttt gctctcacct ctgactccct
      361 acaggctaca gctagagagt tgactcctct atttgaggta agagtgtctg gatttgggat
      421 aaggtgacct cagtttgcag gtctcatggg ggagatcaga gggtcctttg ttgagaagag
      481 gggacttggt ccacagtgag caaaggaatt tgtctaaatt gctattaagg gagatcatct
      541 cccaaggtgg gccaagcctg aaccctcagc cctccatttc ttccctttct tttgccttca
      601 gattgacagg cctcggaagt caaaataagt ggtttcctag accgggtcga gagcaagtct
      661 ctattggtcc caactgagtt ttttcagctg gtttttcaac caaacagcac ctcatctccc
      721 agtgagggga agggaaggct gggctgagag cagcaaggct gctcatctca cctctcccca
      781 cccagccatg ccagccgcca cacctggtgg ggagaggtgg gcctcacctg ggtcccctgg
      841 cagtgctctg tgaagggtct tgacattgca ctgtaataat aaaggtgtgt gtgaagtatc
      901 tttttatggt gactttctaa aacccaggga atcatgggac cagttctgat gactcagcct
      961 ggcttccagt ctcctccagg cccaacggtg gcccccagca ctggttgggg cctgggagag
     1021 ctggccttgg ctgaagtgaa gccacctacc cttcaggcat aacaggacag tgagaaggaa
     1081 ggaaagcctg cctcaacctc ccatcagccc tgagcacccc agaagggggc cggctaggag
     1141 tctaggcatg caggaggctg acccctgact gggctcatat ccagccacaa ggcagccagg
     1201 gacccgggca cccacccctt gtctgcgtcc ctctcgggaa tgggcctctt gcccaggcca
     1261 gaaatacacc acctacagta caaattataa tctaaaaaca agagggtggt gttgagtggg
     1321 gaaattgggg aaggtgtttt aggagccact aggaaaatgg gcagcaggga ctctctggac
     1381 tggcttggga agagcgcttt tggggaacct ggaggatggc aagctgagaa acactggtgt
     1441 ggagattcca gccaaatccc aggcctgccc ctccccctcc tctgagaggc cgtcttcttg
     1501 gcagacagca gagagatgca tgacaaaggt gccgtgatgg ttctgtcctg gggattgaga
     1561 tggctgggga ggggcctcct cctgttccga agcatgttcc tcccaccccc accaggcccc
     1621 ataatctacc tgccttttgg gcagttaaag gccgagaagt gaacacagct gcaaccccac
     1681 tgccttgtag accttccggc agacctgtgg caggtattga aatgcacgca tacaattagg
     1741 ctcaaaaagt ctacacagac aggagatggg cacacgaaca gaggcaacat aagagtgggg
     1801 gaaaagtctc aaaagactca cggatgccac caagatgaag acagctggcc acgggacacc
     1861 catcccctta gaaggcagat agagccactg accccagcag acaagcccag gcagggctga
     1921 gcctggagcc tgcaatgaga agccttactt aagtcgacag aggtcagcgt gcccagtcca
     1981 gacctggcct tctggccttc gaagctgtgg ggagccctgg cccagagccc cctctggagc
     2041 ccccagactt accccaggcc ctccactgag atcaagtttt gggagcagac agacaaacat
     2101 catccctcac agacaggcat tccgttggct attctcttgc aaacagaatc aagcactaga
     2161 ccagcagcat gagcctcagg atactcaggc cgggcccaga aaaacagacc ctgaagggga
     2221 gcttagggca gccttcctgc acccctccac aaatcactct ccacctcctc tgcgtctttc
     2281 tgccagccag ccccactaaa caaagcacat ccctcaatct gccgggctcg gggagggatg
     2341 cacgatgaag ctggacgcct gagtccccca gaggaaggag gaactagata cctaggtccc
     2401 tgtggggggc ccttggtgcc cgtctgaggc tcagtctttg aggggattgc agaggggggt
     2461 tgctggagct ccttttagcg tctctgaagg ggattctgtg tgaggggatt gggactgggg
     2521 ggttggggag caggaagcag tccccagggg agccatccag gcccattcaa gggttgagca
     2581 cttgtttagg gttagagctg ccccctctgg ggaccgggat tgtccagcca aggccattgt
     2641 cctgccccct tcccccagtc cctcccaggc ttctttgaac ctgaagtcag atattttttc
     2701 tccacacccc ccaccccctg gttttcccca cccagggcct agggctggag gcctgggcca
     2761 gggaggtggg ggagggagaa cggggcctac cgtggtatta gatgtctgag ttttggttga
     2821 gaggggagca aggaacctga tgtgcaggtt ccatagtgga gggggcccaa agcgggtgtc
     2881 ttatcactct gtttcagcaa aggttggtaa actgaggccc agtcagtcca aagtctggtc
     2941 ccttgaaggg gaagtaggga ccaacccctt agtctgttag atgaggagag tctggagtct
     3001 gattctggaa gacggagggg tggggggatg gggggtgggg ggatatagca cggaggcctt
     3061 gtctggcagt ctactcttga agatggggtg aaatttggca ggctgggcag atggtgccag
     3121 gcacccaggc tgcggggtgg ctggatttgg ccagtatcgg gatgggaatg cctaggattc
     3181 tggatggatc gggggaaggc ataagggagc agctggccat tgtgcttatg gctgttgatg
     3241 cattgaggga tagcgccaca cacacattca ataaatttga ggagctgaga gggtgactgg
     3301 cccctgaagg cacagtgcca gaggtctgtg gagggggggt caagcacctg ggttcctgaa
     3361 gaacatggag gtgtgggagt gattccagac agctgggatg tgcagagcct gagagagtgc
     3421 cagggagcgg gttgggagtt gaaagttggg tgtggtggct cacgccttta atcatgacac
     3481 tgggcggcag aggcgggagg atttcttgag gacaggaatt caagaccagc ctgggtaaca
     3541 tagcaaggcc ccatctctac taaaaataaa aaaactaaca gggcacagtg gtccaagcct
     3601 gtagtcccag ccacttagga ggctggagca gaaggattgc tttggcccag tagatcgagg
     3661 ctacattgag ccatcattgt actccactgc actccagtct gggcaacaaa gtgagaccct
     3721 gtcttaaaaa ataaaaataa aaaaagtttc tgtgggggac ctgcactgag gtcctggagg
     3781 ggcgccagtt gtgtctcccg gttttcccct tccacagaca ccattgccac caccattagg
     3841 caaacatcct tcgcctcagt ttctcccccc acctccctct cctccaccca tccagggggc
     3901 ggggccagag gtcaaggcta gtgggtggga ctggggaggg agagaggggt tgagtagtcc
     3961 cttcgcaagc cctcatttca ccaggccccc ggcttggggc gccttccttc cccatggcgg
     4021 gacacctggc ttcagatttt gccttctcgg atccaccggt cgccaccatg gtgagcaagg
     4081 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     4141 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     4201 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     4261 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     4321 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     4381 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     4441 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     4501 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     4561 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     4621 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     4681 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     4741 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact
     4801 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     4861 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     4921 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4981 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     5041 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     5101 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5161 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     5221 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5281 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     5341 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     5401 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     5461 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     5521 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5581 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     5641 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     5701 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     5761 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     5821 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5881 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5941 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     6001 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     6061 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     6121 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     6181 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     6241 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     6301 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     6361 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     6421 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     6481 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     6541 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     6601 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     6661 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     6721 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     6781 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     6841 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     6901 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     6961 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     7021 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     7081 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     7141 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     7201 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     7261 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7321 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7381 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     7441 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7501 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7561 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7621 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7681 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7741 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7801 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7861 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7921 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7981 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     8041 ggccttttgc tggccttttg ctcacatg
//
