LOCUS       sequence-244682-        6975 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Addgene next-generation sequencing identified numerous modifications
            from the indicated plasmid backbone. Please see the sequences link 
            for additional information..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6975)
  AUTHORS   Hanna J, Cheng AW, Saha K, Kim J, Lengner CJ, Soldner F, Cassady JP,
            Muffat J, Carey BW, Jaenisch R
  TITLE     Human embryonic stem cells with biological and epigenetic 
            characteristics similar to those of mouse ESCs.
  JOURNAL   Proc Natl Acad Sci U S A. 2010 May 18. 107(20):9222-7.
  PUBMED    20442331
REFERENCE   2  (bases 1 to 6975)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2010 May 18. 107(20):9222-7."
COMMENT     Sequence Label: pGL2-hOct4 DE-SV40-Luc
FEATURES             Location/Qualifiers
     source          1..6975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..1653
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(67..85)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1694..1815
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1705..1724)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1759..1778
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2063..2080)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2234..2822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2314..2333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2993..3784,3785..3853))
                     /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     3616..3635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3854..3958)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3985..4440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4072..4091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4282..4303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4571..4619
                     /note="synthetic polyadenylation signal"
     misc_feature    4633..4724
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4673..4692
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        6744..6940
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6791..6926
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      join(6970..6975,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct ggaagatgga
       61 accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt
      121 gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt cgaaatgtcc
      181 gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta
      241 tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt
      301 gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgggcatt
      361 tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa
      421 aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga ttaccaggga
      481 tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat
      541 tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa ctcctctgga
      601 tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt gagattctcg
      661 catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt
      721 gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt
      781 cgagtcgtct taatgtatag atttgaagaa gagctgtttc tgaggagcct tcaggattac
      841 aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa aagcactctg
      901 attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc tcccctctct
      961 aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag gcaaggatat
     1021 gggctcactg agactacatc agctattctg attacacccg agggggatga taaaccgggc
     1081 gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa
     1141 acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat tatgtccggt
     1201 tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct
     1261 ggagacatag cttactggga cgaagacgaa cacttcttca tcgttgaccg cctgaagtct
     1321 ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat cttgctccaa
     1381 caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc cggtgaactt
     1441 cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat
     1501 tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac
     1561 gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata
     1621 aaggccaaga agggcggaaa gatcgccgtg taattctaga gtcggggcgg ccggccgctt
     1681 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     1741 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     1801 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     1861 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat
     1921 ccgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     1981 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     2041 gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2101 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2161 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2221 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2281 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2341 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2401 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2461 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2521 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2581 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2641 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2701 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2761 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2821 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2881 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2941 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3001 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3061 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3121 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3181 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3241 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3301 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3361 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3421 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3481 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3541 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3601 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3661 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3721 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3781 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3841 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3901 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3961 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4021 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4081 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4141 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4201 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4261 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4321 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4381 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     4441 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     4501 tattacgcca gcccaagcta ccatgataag taagtaatat taaggtacgg gaggtacttg
     4561 gagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     4621 atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa
     4681 ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtacccat
     4741 tgagtccaaa tcctctttac taggtgagga aactgaggca caggaggggg aggtgccatt
     4801 agccagggag tcagagccag gacttagaac ctggtccatg cctcctagtt tgttcctgac
     4861 caatcctccc agctcaccca ctccttttgc tctcacctct gactccctac aggctacagc
     4921 tagagagttg actcctctat ttgaggtaag agtgtctgga tttgggataa ggtgacctca
     4981 gtttgcaggt ctcatggggg agatcagagg gtcctttgtt gagaagaggg gacttggtcc
     5041 acagtgagca aaggaatttg tctaaattgc tattaaggga gatcatctcc caaggtgggc
     5101 caagcctgaa ccctcagccc tccatttctt ccctttcttt tgccttcaga ttgacaggcc
     5161 tcggaagtca aaataagtgg tttcctagac cgggtcgaga gcaagtctct attggtccca
     5221 actgagtttt ttcagctggt ttttcaacca aacagcacct catctcccag tgaggggaag
     5281 ggaaggctgg gctgacagca gcaaggctgc tcatctcacc tctccccacc cagccatgcc
     5341 agccgccaca cctggtgggg agaggtgggc ctcacctggg tcccctggca gtgctctgtg
     5401 aagggtcttg acattgcact gtaataataa aggtgtgtgt gaagtatctt tttatggtga
     5461 ctttctaaaa cccagggaat catgggacca gttctgatga ctcagcctgg cttccagtct
     5521 cctccaggcc caacggtggc ccccagcact ggttggggcc tgggagagct ggccttggct
     5581 gaagtgaagc cacctaccct tcaggcataa caggacagtg agaaggaagg aaagcctgcc
     5641 tcaacctccc atcagccctg agcaccccag aagggggccg gctaggagtc taggcatgca
     5701 ggaggctgac ccctgactgg gctcatatcc agccacaagg cagccaggga cccaggcacc
     5761 caccccttgt ctgcgtccct ctcgggaatg ggcctcttgc ccaggccaga aatacaccac
     5821 ctacagtaca aattataatc taaaaacaag agggtggtgt tgagtgggga aattggggaa
     5881 ggtgttttag gagccactag gaaaatgggc agcagggact ctctggactg gcttgggaag
     5941 agcgcttttg gggaacctgg aggatggcaa gctgagaaac actggtgcgg agattccagc
     6001 caaatcccag gcctgcccct ccccctcctc tgagaggccg tcttcttggc agacagcaga
     6061 gagatgcatg acaaaggtgc cgtgatggtt ctgtcctggg gattgagatg gctggggagg
     6121 ggcctcctcc tgttccgaag catgttcctc ccacccccac caggccccat aatctacctg
     6181 ccttttgggc agttaaaggc cgagaagtga acacagctcc aaccccactg ccttgtagac
     6241 cttccggcag acctgtggca ggtattgaaa tgcacgcata caattaggct caaaaagtct
     6301 acacagacag gagatgggca cacgaacaga ggcaacataa gagtggggga aaagtctcaa
     6361 aagactcacg gatgccacca agatgaagac agctggccac gggacaccca tccccttaga
     6421 aggcagacag agccactgac cccagcagac aagcccaggc agggctgagc ctggagcctg
     6481 caatgagaag ccttacttaa gtcgacagag gtcagcgtgc ccagtccaga cctggccttc
     6541 tggccttcga agctgtgggg agccctggcc cagagccccc tctggagccc ccagacttac
     6601 cccaggccct ccactgagat caagttttgg gagcagacag acaaacatca tccctcacag
     6661 acaggcattc cgttggctat tctcttgcaa acagaatcaa gcactagacc agcagcatga
     6721 gcctcagctc gagatctgcg atctgcatct caattagtca gcaaccatag tcccgcccct
     6781 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatcgctg
     6841 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     6901 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttggca ttccggtact
     6961 gttggtaaag ccacc
//
