LOCUS       sequence-198212-        8842 bp    DNA     circular SYN 14-APR-2025
DEFINITION  in vitro transcription (T7 promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8842)
  AUTHORS   Chung YG, Matoba S, Liu Y, Eum JH, Lu F, Jiang W, Lee JE, Sepilian 
            V, Cha KY, Lee DR, Zhang Y
  TITLE     Histone Demethylase Expression Enhances Human Somatic Cell Nuclear 
            Transfer Efficiency and Promotes Derivation of Pluripotent Stem 
            Cells.
  JOURNAL   Cell Stem Cell. 2015 Dec 3;17(6):758-66. doi: 
            10.1016/j.stem.2015.10.001. Epub 2015 Oct 29.
  PUBMED    26526725
REFERENCE   2  (bases 1 to 8842)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.stem.2015.10.001"; journalName: "Cell Stem Cell"; date: 
            "2015-12-3- 3"; volume: "17"; issue: "6"; pages: "758-66"
COMMENT     Sequence Label: pcDNA-flag-hKDM4A-polyA
FEATURES             Location/Qualifiers
     source          1..8842
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             911..934
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4346..4375
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4391..4408
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4431..4448)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4437..4661
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4707..5135
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4794..4813)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5004..5025
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5144..5164)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5149..5478
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5329..5464
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5391..5410
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5545..6339
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5599..5618)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6209..6228
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6515..6636
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6552..6571)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6606..6625
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6685..6701)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6685..6701)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6698..6720)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6709..6725
                     /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(6733..6739,6740..6757,6758..6763))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6778..6799
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6916..6933)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7087..7675)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7167..7186)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7846..8637,8638..8706))
                     /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     8469..8488
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8707..8811)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttatg gactacaaag acgatgacga caagggatcc atggcttctg agtctgaaac
      961 tctgaatccc agtgctagga taatgacctt ttatccaact atggaagagt tccgaaactt
     1021 cagtagatac attgcctaca ttgaatccca aggagctcat cgggcagggc tagccaaggt
     1081 tgttcctcca aaagagtgga agccacgagc atcctatgat gacattgatg atttggtcat
     1141 tcctgccccc attcaacagc tggtgacggg gcagtctggc ctctttactc agtacaacat
     1201 acagaagaaa gccatgactg ttcgagagtt ccgcaagata gccaatagcg ataagtactg
     1261 taccccacgc tatagtgagt ttgaagagct cgagcggaaa tactggaaaa atcttacatt
     1321 caatcctcca atctatggtg cagatgtgaa tggtaccctc tatgaaaagc atgttgatga
     1381 gtggaatatt ggccggctga gaacaatcct ggacttggtg gaaaaggaga gtgggatcac
     1441 cattgagggt gtgaacaccc catacctgta ctttggcatg tggaagacat cctttgcttg
     1501 gcacactgaa gacatggacc tctacagcat caactacctg cactttggag aaccaaagtc
     1561 ctggtactct gttccacctg agcatggaaa gcggttggaa cgcctcgcca aaggcttttt
     1621 cccaggaagt gctcaaagct gtgaggcatt tctccgccac aagatgaccc tgatttcccc
     1681 gttaatgctg aagaaatatg gaattccctt tgacaaggtg actcaagagg ctggagagtt
     1741 tatgatcact ttcccttatg gttaccatgc cggctttaac catggtttta actgtgcgga
     1801 gtctaccaat tttgctaccc gtcggtggat tgagtacggc aagcaagctg tgctgtgctc
     1861 ctgtagaaag gacatggtga agatctccat ggatgtgttt gtgagaaagt tccagccaga
     1921 aaggtacaaa ctttggaaag ctgggaagga caacacagtt attgaccata ctctgcccac
     1981 gccagaagca gctgagtttc ttaaggagag tgaactgcct ccaagagctg gcaacgagga
     2041 ggagtgccca gaggaggaca tggaaggggt ggaggatgga gaggaaggag acctgaagac
     2101 aagcctggcc aagcaccgaa tagggacaaa gaggcaccga gtttgtcttg aaataccaca
     2161 ggaggtgagt cagagtgagc tcttccccaa ggaggatctg agttctgagc agtatgagat
     2221 gacggagtgc ccggcagccc tcgcccctgt gaggcccacc catagctctg tgcggcaagt
     2281 tgaggatggt cttaccttcc cagattattc tgactccact gaagtcaaat ttgaagagct
     2341 taaaaatgtc aaactagaag aggaggatga ggaggaagaa caagaagcag ctgccttgga
     2401 tctttctgtg aatcctgcgt ctgtaggggg acgccttgtc ttctcaggct ccaaaaagaa
     2461 atcatcttct agcctgggct ctggctcttc acgggattct atctcttctg attcagaaac
     2521 tagtgagcct ctctcctgcc gagcccaagg gcaaacggga gttctcactg tgcacagtta
     2581 tgccaaaggg gatggcaggg tcactgtggg agagccatgc acgaggaaga aaggaagcgc
     2641 cgctagaagt ttcagtgagc gggagctggc agaggttgca gatgaataca tgttttccct
     2701 agaagagaat aagaagtcca agggacgccg tcagccttta agcaagctcc cccgccatca
     2761 cccacttgtg ctgcaggagt gtgtcagtga tgatgagaca tctgaacagc tgacccctga
     2821 ggaagaggct gaggagacag aggcctgggc caagcctctg agccaactgt ggcagaaccg
     2881 acctccaaac tttgaggctg agaaggaatt caatgagacc atggcccaac aggcccctca
     2941 ctgcgctgtc tgtatgatct tccagactta tcatcaggtt gaatttggag gctttaatca
     3001 gaactgtgga aatgcttcag atttagcccc ccagaagcag aggaccaagc cattgattcc
     3061 agaaatgtgc ttcacttcga ctggctgcag cacggacatc aacctttcta ctccttatct
     3121 tgaggaggat ggcaccagca tactcgtttc ctgcaagaag tgcagcgtcc gggtccatgc
     3181 cagttgctat ggggtccccc ctgcaaaggc ttctgaagac tggatgtgtt ctcggtgttc
     3241 agccaatgcc ctagaggagg actgctgttt atgctcatta cgaggagggg ccctgcagag
     3301 agcaaatgat gacaggtggg tccacgtttc atgtgctgtg gcaattctgg aagcaaggtt
     3361 tgtcaacatt gcagaaagaa gtccggtgga tgtgagcaaa atccccctgc cccgcttcaa
     3421 actgaaatgt atcttctgta agaagcggag gaaaagaact gctggctgct gtgtgcagtg
     3481 ttctcacggc cgctgcccaa ctgccttcca tgtgagctgc gcccaggctg ccggtgtgat
     3541 gatgcagcct gacgactggc cttttgtggt cttcattacc tgctttcggc acaagattcc
     3601 taatttggag cgtgccaagg gggccttgca aagcatcact gcaggccaga aagtcattag
     3661 caagcataag aacgggcgct tctaccagtg tgaagtggtc aggctcacca ccgagacctt
     3721 ctatgaagtc aactttgatg atggctcctt cagcgacaat ctttatcctg aggacatagt
     3781 gagccaggac tgtctccagt ttggtcctcc tgctgaaggg gaagtggtcc aagtgagatg
     3841 gacagacggc caagtctatg gagccaagtt tgtggcctcc caccctatcc aaatgtacca
     3901 ggtggagttt gaggatggct cacaacttgt ggttaagaga gatgatgtat acacactgga
     3961 tgaagagctt cccaagagag tcaaatctag actgtcagta gcctcagaca tgcgcttcaa
     4021 tgagattttc acagagaaag aggttaagca agaaaagaaa cggcaacgag ttatcaactc
     4081 aagataccgg gaagattata ttgagcctgc actataccgg gccatcatgg agtagcggcc
     4141 gctcgactat gggcaccaaa gaacctgtaa acgttatctt tttaaattga atgtgcacaa
     4201 ataaaagttt ggaaaagaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4261 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa tacgcatcat acgctgtggc
     4321 gtacctcgag tctagagggc ccttcgaaca aaaactcatc tcagaagagg atctgaatat
     4381 gcataccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     4441 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     4501 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     4561 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4621 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     4681 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4741 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4801 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4861 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4921 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4981 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5041 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5101 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     5161 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5221 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5281 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     5341 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5401 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5461 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     5521 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5581 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5641 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5701 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5761 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5821 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5881 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5941 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     6001 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     6061 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     6121 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     6181 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     6241 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6301 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     6361 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6421 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6481 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     6541 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6601 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     6661 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6721 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6781 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6841 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6901 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6961 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7021 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7081 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7141 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7201 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7261 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7321 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7381 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7441 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7501 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7561 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7621 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7681 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7741 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7801 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7861 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7921 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7981 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8041 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8101 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8161 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8221 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8281 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8341 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8401 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8461 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8521 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8581 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8641 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8701 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8761 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8821 ccgaaaagtg ccacctgacg tc
//
