LOCUS       sequence-315972-        6812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Marmoset Targeting.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6812)
  AUTHORS   Yoshimatsu S, Murakami R, Sato T, Saeki T, Yamamoto M, Sasaki E, 
            Noce T, Okano H
  TITLE     Generation of a common marmoset embryonic stem cell line CMES40-OC 
            harboring a POU5F1 (OCT4)-2A-mCerulean3 knock-in reporter allele.
  JOURNAL   Stem Cell Res. 2021 Mar 24;53:102308. doi: 
            10.1016/j.scr.2021.102308.
  PUBMED    33799281
REFERENCE   2  (bases 1 to 6812)
  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"; journalName: "Stem Cell 
            Res."; date: "2021-03-24"; volume: "53:102308. doi"; pages: " 
            10.1016/j.scr.2021.102308"
COMMENT     Sequence Label: pKI-cjOCT4-Cyan-pac
FEATURES             Location/Qualifiers
     source          1..6812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     186..208
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     200..217
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     201..217
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     224..243
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        224..242
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1399..1455
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(1456..1458,1459..1461,1462..2172)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGNVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1762..1781)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2109..2130
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2179..2232
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2233..2832
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2233..2252)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2729..2749
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     promoter        complement(3686..3704)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3687..3704)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3722..3738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3722..3738)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3735..3757)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3746..3762
                     /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(3770..3776,3777..3794,3795..3800))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3815..3836
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3953..3970)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4124..4712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4204..4223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4850..5224)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      5221..5230
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5430..6224)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(5430..6224)
                     /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(5541..5560)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6151..6170
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(join(6573..6812,1..63))
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(6675..6694)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 actctgtaca tccacaaaca gacgataacg gctctctctt ttataggtgt aaaccttaaa
       61 ctgccgtacg tataggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
      121 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      181 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata
      241 gggcgaattg ggccctctag atgcatgctc gagcggccgc cagtgtgatg gatatctgca
      301 gaattcgccc ttagaggtgg gaagtctgga tcttggagtc aacaggggcc tgcgtgttcc
      361 ctcattttgc ttctcttttg acttgcctcc cccaggaaag gtgttcagcc aaacgaccat
      421 ctgccgcttt gaggctctgc agcttagctt caagaacatg tgtaagctgc ggcctttact
      481 gcagaagtgg gtggaggaag ctgacaacaa tgaaaatctt caggaggtaa gggtgggagg
      541 gggatacacg aggaccttcc ctttcttggc ccagtttcca ttgcttccat tcctggcttg
      601 tggctctccc tctttggtgc agttctcagt gagatggagt aaaatgcctc ccttctgctt
      661 ggagaaggtc cagagtcaac ctttccagga tcctgccttt acacaccacc tggctctgct
      721 gacacatcta gtcacagacc ccggtgacgc tgttactcag caagtccaaa gcttgccctt
      781 gtaccccctt cccacctgca cagatatgca aagcagaaac cctcgtgcag gcccggaaga
      841 gaaagcgaac cagtatcgag aatcgagtga gaggcagcct ggagaatttg ttcctgcagt
      901 gcccgaaacc cacacttcag cagatcagcc acatcgccca gcagcttggg ctcgagaagg
      961 atgtgagtgg cgagcctctc catctctttc ctctgtcacc acctctgctt tctctagctc
     1021 tggctcctcc tccactctag ggctgttggc tttgggcaga atgacccggt agggtcaggc
     1081 ccgtctctcc gttcattgtc taatgtcact ctcctctgtg ttgtccactc gcaggtggtc
     1141 cgtgtgtggt tctgtaaccg gcgccagaag ggcaagcgat caagcagtga ctatgcacaa
     1201 cgagaggatt ttgaggctgc tgggtcccct ttctcagggg gaccagtgtc ctttcctctg
     1261 gccccagggc cccattttgg taccccaggc tatgggagcc ctcatttcac tgcactgtac
     1321 tcctcggtcc cattccctga gggggaagcc ttcccccctg tctctgtccc cactctgggc
     1381 tctcccatgc attccaacgc cacgaacttc tctctgttaa agcaagcagg agacgtggaa
     1441 gaaaaccccg gtcctatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1501 ctggtcgagc tggacggcaa cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1561 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1621 gtgccctggc ccaccctcgt gaccaccctg agctggggcg tgcagtgctt cgcccgctac
     1681 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1741 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1801 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1861 aacatcctgg ggcacaagct ggagtacaac gccatccacg gcaacgtcta tatcaccgcc
     1921 gacaagcaga agaacggcat caaggccaac ttcggcctca actgcaacat cgaggacggc
     1981 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2041 ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag
     2101 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2161 gagctgtaca aggtcgacga gggcagagga agtcttctaa catgcgggga cgtggaggaa
     2221 aatcccggcc ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     2281 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     2341 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     2401 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     2461 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     2521 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     2581 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     2641 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     2701 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     2761 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     2821 cccggtgcct gaggtgccta tcctaggaat gggggacagg gagaggagaa gctggggaaa
     2881 gagaacctgg agtttgtgcc agggctttta ggattaagtt cttcattcac taaggaaggg
     2941 attgggaaca caaagggtga gggcagggga gtttagggca actggttaga gggaaggtga
     3001 aattcaatga tgctcttgat tttaatcccc acatgtatca cttttttctt aaataaagaa
     3061 gcctgggaca cagtagatag acacatttat cttggtttgg cctcctttag ttactgaggt
     3121 ggggatggga atatccaatg ctcataccca agtgacccta aaaccaagat gccatttaaa
     3181 ctccttgagg tcacacaatg tcagaggaag agctaggatt ggagccaagt ttatctgcat
     3241 aggatccctt tatagctagt tcatggaagc ctgtgacatg ggatctggac accacacttt
     3301 tgcccatatt catcaccatg tagctgttac ggctcagagc tggctttttg ctttccccac
     3361 cctactctgg aattcttaaa tggctttata cttataatca agtcttattt ctgttgtacc
     3421 taaatcaccc caactggaaa cttctattaa tattttgtgc tttcttggta ccagggtcta
     3481 tctggtttcc acttaaggtt tttgcatacc ctgcccataa gtgactcacc aaatacataa
     3541 ggctttattt ctgtacaaat gatgctttga caatagcttg gcttggtgta aggctcacaa
     3601 aggaaaaagg gcgaattcca gcacactggc ggccgttact agtggatccg agctcggtac
     3661 caagcttgat gcatagcttg agtattctat agtgtcacct aaatagcttg gcgtaatcat
     3721 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3781 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3841 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3901 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3961 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4021 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4081 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4141 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4201 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4261 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4321 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4381 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4441 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4501 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4561 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4621 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4681 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4741 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4801 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt tagcacgtgt cagtcctgct
     4861 cctcggccac gaagtgcacg cagttgccgg ccgggtcgcg cagggcgaac tcccgccccc
     4921 acggctgctc gccgatctcg gtcatggccg gcccggaggc gtcccggaag ttcgtggaca
     4981 cgacctccga ccactcggcg tacagctcgt ccaggccgcg cacccacacc caggccaggg
     5041 tgttgtccgg caccacctgg tcctggaccg cgctgatgaa cagggtcacg tcgtcccgga
     5101 ccacaccggc gaagtcgtcc tccacgaagt cccgggagaa cccgagccgg tcggtccaga
     5161 actcgaccgc tccggcgacg tcgcgcgcgg tgagcaccgg aacggcactg gtcaacttgg
     5221 ccatggtggc cctcctcacg tgctattatt gaagcattta tcagggttat tgtctcatga
     5281 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5341 cccgaaaagt gccacctgat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     5401 cgcatcagga aattgtaagc gttaataatt cagaagaact cgtcaagaag gcgatagaag
     5461 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     5521 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
     5581 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     5641 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgctcgcc
     5701 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     5761 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     5821 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     5881 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     5941 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     6001 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca
     6061 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     6121 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     6181 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
     6241 cctgtctctt gatcagagct tgatcccctg cgccatcaga tccttggcgg caagaaagcc
     6301 atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc
     6361 ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa
     6421 gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca
     6481 ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtgaaaag gatctaggtg
     6541 aagatccttt ttgataatct catgcctgac atttatattc cccagaacat caggttaatg
     6601 gcgtttttga tgtcattttc gcggtggctg agatcagcca cttcttcccc gataacggag
     6661 accggcacac tggccatatc ggtggtcatc atgcgccagc tttcatcccc gatatgcacc
     6721 accgggtaaa gttcacggga gactttatct gacagcagac gtgcactggc cagggggatc
     6781 accatccgtc gccccggcgt gtcaataata tc
//
