LOCUS       sequence-101002-        7226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mRNA synthesis or expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7226)
  AUTHORS   Matoba S, Liu Y, Lu F, Iwabuchi KA, Shen L, Inoue A, Zhang Y
  TITLE     Embryonic Development following Somatic Cell Nuclear Transfer 
            Impeded by Persisting Histone Methylation.
  JOURNAL   Cell. 2014 Nov 6;159(4):884-95. doi: 10.1016/j.cell.2014.09.055. 
            Epub 2014 Oct 30.
  PUBMED    25417163
REFERENCE   2  (bases 1 to 7226)
  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: 
            "10.1016/j.cell.2014.09.055"; journalName: "Cell"; date: "2014-11-6-
            6"; volume: "159"; issue: "4"; pages: "884-95"
COMMENT     Sequence Label: pcDNA-flag-mZscan4d-polyA
FEATURES             Location/Qualifiers
     source          1..7226
                     /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             2729..2758
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2774..2791
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2814..2831)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2820..3044
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3090..3518
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3177..3196)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3387..3408
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3527..3547)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3532..3862
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3713..3848
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3775..3794
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3929..4723
                     /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(3983..4002)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4593..4612
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4899..5020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4936..4955)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4990..5009
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5069..5085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5069..5085)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5082..5104)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5093..5109
                     /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(5117..5123,5124..5141,5142..5147))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5162..5183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5300..5317)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5471..6059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5551..5570)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6230..7021,7022..7090))
                     /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     6853..6872
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7091..7195)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 actagtaacg gccgccagtg
      961 tgctggaatt cacgcttcac agcaggcacc agcaaaagac cttcagacca acaatttaga
     1021 gtttactcca tctcatagtt ctggtgtgca gtgggtagaa gacatctcta actcaccaag
     1081 tgctcagcta aacttttctc caagtaacaa tggctgctgg gcaactcagg agctgcaaag
     1141 tctctggaag atgttcaact cctggttgca gccagaaaag cagactaagg agcagatgat
     1201 ttctcaactg gtcttggagc agtttctcct cattgggcac tgcaaggaca agtatgcttt
     1261 gacagagaag tggaaagcca gtggtagcga tatgaggaga ttcatggaga gtctgactga
     1321 tgagtgcttg aagcctcctg tcatggtcca tgtttcaatg caaggacaag aagctctctt
     1381 ttctgaaaac atgccattaa aagaagtcat caagcttttg aaacaacagc aatctgcaac
     1441 aaggccaaca ccagataatg agcagatgcc agtagacacc acacaagata gattattggc
     1501 cacaggacaa gaaaacagtg aaaatgaatg caacaactct tgtaatgcta ctgaagcaaa
     1561 tgttggtgaa agctgtagtg gaaatgaaat ggactccctt cttattatcc agaaagaaca
     1621 gtaccctgag catgaagagg ggaatgttgt ttttcaattc cctcttgatg ccagaagagc
     1681 aagtcaaggc aactccagtc atcatgtaga cttccggagt gctccgactc ctgcggatgt
     1741 ccccatggag gaacaaccaa aggatttatc cagagaaaac atctctgagg acaagaacaa
     1801 ttgctataac acttccagga atgcagctac tcaagtatat agaagtgata atattcccag
     1861 gaaaaagaca gactcccttt ccattaacaa gagaatatat cattctgagc ctgaggaggg
     1921 agatattcct tatggagttc ctcaggattc tacaagagca agtcaaggaa catctacatg
     1981 cttgcaagag tcacttgggg aatgtttttc tgaaaaagac cctagggagc taccagggtt
     2041 ggagtctagg caagaggagc ctatctctga tcctgtcttt cttggtaagg atcatgaggc
     2101 aaacttacca tgtgaaagtc atcaaaagag attccgtaga gatgccaaac tattcaagtg
     2161 tgaagaatgt tctaggatgt tcaaacatgc caggagcctt tcgtcccacc agagaactca
     2221 cctgaataag aagagtgaat tgctttgtgt cacctgtcag aaaatgttca aacgagtctc
     2281 tgaccgccga acccatgaga tcatacacat gccagaaaag cctttcaagt gcagcacatg
     2341 tgaaaagtcc ttcagccaca agaccaacct gaagtctcat gagatgattc acacaggaga
     2401 aatgccttat gtctgttccc tatgtagccg tcgctttcgc caatcatcca cttaccatcg
     2461 tcacctgagg aattaccaca gatctgacta agaattctgc agatatccat cacactggcg
     2521 gccgctcgac tatgggcacc aaagaacctg taaacgttat ctttttaaat tgaatgtgca
     2581 caaataaaag tttggaaaag aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2641 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaatacgcat catacgctgt
     2701 ggcgtacctc gagtctagag ggcccttcga acaaaaactc atctcagaag aggatctgaa
     2761 tatgcatacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     2821 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2881 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2941 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3001 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3061 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3121 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3181 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3241 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3301 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3361 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3421 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     3481 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3541 ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa gcatgcatct
     3601 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3661 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3721 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3781 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3841 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3901 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3961 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4021 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4081 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4141 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4201 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4261 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4321 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4381 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4441 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4501 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4561 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4621 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4681 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4741 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4801 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4861 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4921 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4981 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5041 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5101 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5161 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5221 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5281 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5341 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5401 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5461 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5521 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5581 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5641 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     5701 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5761 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5821 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5881 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     5941 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6001 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6061 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6121 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6181 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6241 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6301 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6361 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6421 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6481 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6541 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6601 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6661 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6721 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6781 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6841 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6901 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6961 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7021 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7081 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7141 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7201 ttccccgaaa agtgccacct gacgtc
//
