LOCUS       Exported                8068 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    phOct4-EGFP
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8068
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      4008..4017
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4014..4733
                     /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             4014..4730
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4059..4080)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4320..4339)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4667..4688
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4856..4977
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4893..4912)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4947..4966
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4984..5439)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5121..5142)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5333..5352
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5466..5570
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5572..5929
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(5595..5615)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5780..5915
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5842..5861
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5964..6758
                     /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(6018..6037)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6628..6647
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6946..6965)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6990..7037
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7366..7954
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7855..7874
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattac tagcgctacc ggactcagat ctcgagctca agcttcccat gtcaagtaag
       61 tggggtgggt agatgggagg gcatatgaga gcggggcagc aattttatgt cttagttgga
      121 aggaagttta gagttcattg agtccaaatc ctctttacta ggtgaggaaa ctgaggcaca
      181 ggagggggag gtgccattag ccagggagtc agagccagga cttagaacct ggtccatgcc
      241 tcctagtttg ttcctgacca atcctcccag ctcacccact ccttttgctc tcacctctga
      301 ctccctacag gctacagcta gagagttgac tcctctattt gaggtaagag tgtctggatt
      361 tgggataagg tgacctcagt ttgcaggtct catgggggag atcagagggt cctttgttga
      421 gaagagggga cttggtccac agtgagcaaa ggaatttgtc taaattgcta ttaagggaga
      481 tcatctccca aggtgggcca agcctgaacc ctcagccctc catttcttcc ctttcttttg
      541 ccttcagatt gacaggcctc ggaagtcaaa ataagtggtt tcctagaccg ggtcgagagc
      601 aagtctctat tggtcccaac tgagtttttt cagctggttt ttcaaccaaa cagcacctca
      661 tctcccagtg aggggaaggg aaggctgggc tgagagcagc aaggctgctc atctcacctc
      721 tccccaccca gccatgccag ccgccacacc tggtggggag aggtgggcct cacctgggtc
      781 ccctggcagt gctctgtgaa gggtcttgac attgcactgt aataataaag gtgtgtgtga
      841 agtatctttt tatggtgact ttctaaaacc cagggaatca tgggaccagt tctgatgact
      901 cagcctggct tccagtctcc tccaggccca acggtggccc ccagcactgg ttggggcctg
      961 ggagagctgg ccttggctga agtgaagcca cctacccttc aggcataaca ggacagtgag
     1021 aaggaaggaa agcctgcctc aacctcccat cagccctgag caccccagaa gggggccggc
     1081 taggagtcta ggcatgcagg aggctgaccc ctgactgggc tcatatccag ccacaaggca
     1141 gccagggacc caggcaccca ccccttgtct gcgtccctct cgggaatggg cctcttgccc
     1201 aggccagaaa tacaccacct acagtacaaa ttataatcta aaaacaagag ggtggtgttg
     1261 agtggggaaa ttggggaagg tgttttagga gccactagga aaatgggcag cagggactct
     1321 ctggactggc ttgggaagag cgcttttggg gaacctggag gatggcaagc tgagaaacac
     1381 tggtgtggag attccagcca aatcccaggc ctgcccctcc ccctcctctg agaggccgtc
     1441 ttcttggcag acagcagaga gatgcatgac aaaggtgccg tgatggttct gtcctgggga
     1501 ttgagatggc tggggagggg cctcctcctg ttccgaagca tgttcctccc acccccacca
     1561 ggccccataa tctacctgcc ttttgggcag ttaaaggccg agaagtgaac acagctgcaa
     1621 ccccactgcc ttgtagacct tccggcagac ctgtggcagg tattgaaatg cacgcataca
     1681 attaggctca aaaagtctac acagacagga gatgggcaca cgaacagagg caacataaga
     1741 gtgggggaaa agtctcaaaa gactcacgga tgccaccaag atgaagacag ctggccacgg
     1801 gacacccatc cccttagaag gcagacagag ccactgaccc cagcagacaa gcccaggcag
     1861 ggctgagcct ggagcctgca atgagaagcc ttacttaagt cgacagaggt cagcgtgccc
     1921 agtccagacc tggccttctg gccttcgaag ctgtggggag ccctggccca gagccccctc
     1981 tggagccccc agacttaccc caggccctcc actgagatca agttttggga gcagacagac
     2041 aaacatcatc cctcacagac aggcattccg ttggctattc tcttgcaaac agaatcaagc
     2101 actagaccag cagcatgagc ctcaggatac tcaggccagg cccagaaaaa cagaccctga
     2161 aggggagctt agggcagcct tcctgcaccc ctccacaaat cactcgccac ctcctctgcg
     2221 tctttctgcc agccagcccc actaaacaaa gcacatccct caatcttccg ggctcgggga
     2281 gggatgcacg atgaagctgg acgcctgagt cccccagagg aaggaggaac tagataccta
     2341 ggtccctgta gggggccctt ggtgcccgtc tgaggctcag tctttgaggg gattgcagag
     2401 gggggttgct ggagctcctt ttagcgtctc tgaaggggat tctgtgtgag gggattggga
     2461 ctggggggtt ggggagcagg aagcagtccc caggggagcc atccaggccc attcaagggt
     2521 tgagcacttg tttagggtta gagctgcccc ctctggggac cgggattgtc cagccaaggc
     2581 cattgtcctg cccccttccc ccagtccctc ccaggcttct ttgaacctga agtcagatat
     2641 tttttctcca caccccccac cccctggttt tccccaccca gggcctaggg ctggaggcct
     2701 gggccaggga ggtgggggag ggagaacggg gcctaccgtg gtattagatg tctgagtttt
     2761 ggttgagagg ggagcaagga acctgatgtg caggttccat agtggagggg gcccaaagcg
     2821 ggtgtcttat cactctgttt cagcaaaggt tgggaaactg aggcccaatc agtccaaagt
     2881 ctggtccctt gaaggggaag taggggccaa ccccttagtc tgttagatga ggagagtctg
     2941 gagtctgatt ctggaagacg gaggggtggg gggatggggg gtggggggat atagcacgga
     3001 ggccttgtct ggcagtctac tcttgaagat ggggtgaaat ttggcaggct gggcagatgg
     3061 tgccaggcac ccaggctgcg gggtggctgg atttggccag tatcgggatg ggaatgccta
     3121 ggattctgga tggatcgggg gaaggcataa gggagcagct ggccattgtg cttatggctg
     3181 ttgatgcatt gagggatagc gccacacaca cattcaataa atttgaggag ctgagagggt
     3241 gactggcccc tgaaggcaca gtgccagagg tctgtggaga gggggtcaag cacctgggtt
     3301 cctgaagaac atggaggtgt gggagtgatt ccagacagct gggatgtgca gagcctgaga
     3361 gagtgccagg gagcgggttg ggagttgaaa gttgggtgtg gtggctcacg cctttaatca
     3421 tgacactggg cggcagaggc gggaggattt cttgaggaca ggaattcaag accagcctgg
     3481 gtaacatagc aaggccccat ctctactaaa aataaaaaaa ctaacagggc acagtggtcc
     3541 aagcctgtag tcccagccac ttaggaggct ggagcagaag gattgctttg gcccagtaga
     3601 tcgaggctac attgagccat cattgtactc cactgcactc cagtctgggc aacaaagtga
     3661 gaccctgtct taaaaaataa aaataaaaaa agtttctgtg ggggacctgc actgaggtcc
     3721 tggaggggcg ccagttgtgt ctcccggttt tccccttcca cagacaccat tgccaccacc
     3781 attaggcaaa catccttcgc ctcagtttct ccccccacct ccctctcctc cacccatcca
     3841 gggggcgggg ccagaggtca aggctagtgg gtgggactgg ggagggagag aggggttgag
     3901 tagtcccttc gcaagccctc atttcaccag gcccccggct tggggcgcct tccttcccca
     3961 tggcgggaca cctggcttca gattttgcct tctcggatcc accggtcgcc accatggtga
     4021 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     4081 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     4141 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     4201 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     4261 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     4321 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     4381 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     4441 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     4501 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     4561 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     4621 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     4681 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc
     4741 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     4801 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     4861 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4921 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     4981 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     5041 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5101 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     5161 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     5221 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     5281 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     5341 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     5401 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     5461 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5521 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     5581 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     5641 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     5701 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     5761 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5821 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     5881 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     5941 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     6001 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     6061 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     6121 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     6181 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     6241 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     6301 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     6361 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     6421 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     6481 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     6541 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     6601 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     6661 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     6721 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     6781 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     6841 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6901 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     6961 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     7021 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     7081 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     7141 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     7201 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7261 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7321 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7381 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     7441 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     7501 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     7561 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     7621 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     7681 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     7741 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     7801 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     7861 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     7921 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     7981 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     8041 tgtggataac cgtattaccg ccatgcat
//
