LOCUS       sequence-36610-e        6102 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 6102)
  AUTHORS   Pasque V, Gillich A, Garrett N, Gurdon JB
  TITLE     Histone variant macroH2A confers resistance to nuclear 
            reprogramming.
  JOURNAL   EMBO J. 2011 May 6;30(12):2373-87. doi: 10.1038/emboj.2011.144.
  PUBMED    21552206
REFERENCE   2  (bases 1 to 6102)
  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: 
            "10.1038/emboj.2011"; journalName: "EMBO J"; date: "2011-05-6- 6"; 
            volume: "30"; issue: "12"; pages: "2373-87"
COMMENT     Sequence Label: pCS2+ macroH2A1.2-GFP-HA
FEATURES             Location/Qualifiers
     source          1..6102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1173..1197)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1205..1207,1208..1210,1211..1921)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1205..1207,1208..1210,1211..1921)
                     /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(1250..1271)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1511..1530)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1858..1879
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1928..2008
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     polyA_signal    2039..2173
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2063..2082)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2117..2136
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2286..2306)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2286..2304)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2325..2341)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2325..2341)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2338..2360)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2349..2365
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2373..2379,2380..2397,2398..2403))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2418..2439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2556..2573)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2727..3315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2807..2826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3486..4277,4278..4346))
                     /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     4109..4128
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4347..4451)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4477..4932)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4614..4635)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4826..4845
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5006..5990
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        6025..6043
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6025..6042
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    join(6098..6102,1..20)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 tttgtacaaa aaagcaggct ccgcggccgc ccccttcacc atgtcgagcc gcggcgggaa
       61 gaagaaatcc accaagacct cccggtcagc caaggccgga gtcatcttcc ctgtgggacg
      121 catgcttcgg tacatcaaga aaggccaccc taagtatagg atcggagtgg gggcacctgt
      181 gtacatggct gctgtcctgg agtacctgac tgctgagatc ctggagctgg ctggcaatgc
      241 agcaagagac aacaagaagg gacgggtcac accccggcac atcctgttag ctgtggccaa
      301 tgatgaagag ctaaaccagc tgctaaaggg tgtcaccata gccagcgggg gcgtgttgcc
      361 gaatatccat cctgagttgc tagcgaagaa gcgaggatcc aagggaaaat tggaagccat
      421 catcacgcct ccgccggcca aaaaggccaa gtctccatcc cagaagaagc cagtggctaa
      481 gaagacagga ggcaagaaag gggcccggaa gtctaagaag aagcagggag aagtgagcaa
      541 ggcggccagc gcagacagta cgacggaggg cacgcctaca gacggcttca ctgtcctctc
      601 caccaagagc ctcttcctcg gccagaagct gaaccttatt cacagtgaaa tcagtaattt
      661 agccggcttt gaggtggagg ccataatcaa tcctaccaat gctgacattg accttaaaga
      721 tgacctagga aacacactgg agaagaaggg cggcaaggag tttgtagaag ctgttctgga
      781 actccggaaa aagaacgggc ccttggaggt agctggagct gctattagtg caggccatgg
      841 cctgcctgcc aagtttgtga tccactgtaa tagtcctgtc tggggtgcag acaaatgtga
      901 agaacttcta gaaaagacgg tgaaaaactg cttggctcta gctgatgaca gaaagctgaa
      961 atccatcgcc ttcccatcca ttggcagcgg caggaacggg ttcccgaagc agacagcggc
     1021 ccagctcatt ctgaaggcca tctccagcta ctttgtctcc acgatgtcct cctccatcaa
     1081 aactgtgtac ttcatgcttt ttgacagtga gagcataggt atctatgtgc aggaaatggc
     1141 caagctggac gccaacaagg gtgggcgcgc cgacccagct ttcttgtaca aagtggttgc
     1201 tagcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1261 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1321 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1381 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1441 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1501 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1561 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1621 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1681 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1741 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1801 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1861 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1921 gctcgagtac ccatacgatg ttccagatta cgcttatcct tatgacgtac ctgactatgc
     1981 atacccttat gatgtaccag actacgctat ctagaactat agtgagtcgt attacgtaga
     2041 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2101 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2161 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2221 gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt
     2281 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     2341 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2401 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2461 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2521 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2581 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2641 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2701 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2761 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2821 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2881 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2941 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3001 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3061 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3121 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     3181 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3241 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3301 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3361 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3421 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3481 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3541 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3601 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3661 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3721 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3781 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3841 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3901 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3961 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4021 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4081 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4141 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4201 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4261 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4321 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4381 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4441 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     4501 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4561 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4621 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4681 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4741 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4801 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4861 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4921 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4981 gggcctcttc gctattacgc cagtcgacca tagccaattc aatatggcgt atatggactc
     5041 atgccaattc aatatggtgg atctggacct gtgccaattc aatatggcgt atatggactc
     5101 gtgccaattc aatatggtgg atctggaccc cagccaattc aatatggcgg acttggcacc
     5161 atgccaattc aatatggcgg acttggcact gtgccaactg gggaggggtc tacttggcac
     5221 ggtgccaagt ttgaggaggg gtcttggccc tgtgccaagt ccgccatatt gaattggcat
     5281 ggtgccaata atggcggcca tattggctat atgccaggat caatatatag gcaatatcca
     5341 atatggccct atgccaatat ggctattggc caggttcaat actatgtatt ggccctatgc
     5401 catatagtat tccatatatg ggttttccta ttgacgtaga tagcccctcc caatgggcgg
     5461 tcccatatac catatatggg gcttcctaat accgcccata gccactcccc cattgacgtc
     5521 aatggtctct atatatggtc tttcctattg acgtcatatg ggcggtccta ttgacgtata
     5581 tggcgcctcc cccattgacg tcaattacgg taaatggccc gcctggctca atgcccattg
     5641 acgtcaatag gaccacccac cattgacgtc aatgggatgg ctcattgccc attcatatcc
     5701 gttctcacgc cccctattga cgtcaatgac ggtaaatggc ccacttggca gtacatcaat
     5761 atctattaat agtaacttgg caagtacatt actattggaa ggacgccagg gtacattggc
     5821 agtactccca ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa gtacatcccc
     5881 attgacgtca atggggaggg gcaatgacgc aaatgggcgt tccattgacg taaatgggcg
     5941 gtaggcgtgc ctaatgggag gtctatataa gcaatgctcg tttagggaac cgccattctg
     6001 cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag ctacttgttc
     6061 tttttgcagg atcccatcga ttcgaattca aggatcaaca ag
//
