LOCUS       sequence-315975-        8175 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 8175)
  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 8175)
  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-cjVASA-tdTomato
FEATURES             Location/Qualifiers
     source          1..8175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..256
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1899..1955
                     /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             1956..3383
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMN
                     FEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVL
                     KGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYER
                     SEGRHHLFLYGMDELYK"
     primer_bind     complement(5063..5082)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5064..5082)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5089..5106)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5089..5105)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5098..5120)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             5243..5545
                     /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     5424..5443
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5894..6688
                     /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             5894..6688
                     /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(5948..5967)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6558..6577
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      6888..6897
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6894..7268
                     /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"
     rep_origin      7406..7994
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7895..7914
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8148..8165
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gcccttagct gccagagggc tagatattga
      361 aaatgtacag catgttatca attttgatct tccttctacc attgatgaat atgttcatcg
      421 aattgggcgt actggtcgtt gtgggaatac tggcagagca atttcctttt ttgatcctga
      481 atcggataac catttagcac agcctctaat aaaagtatta acagatgtaa gttaaacttt
      541 taaaacagaa tggatagttt tcctttgttg ttgaaagtag acattttatg tatatgtata
      601 taacaagtaa acttttccaa tctttaagac tttatgttgc atatgaagtc attttttttt
      661 ttaaggctca acaggatgtt cctgcatggc tggaagaaat tgcctttagt gcatacattc
      721 ctggcttcag tggtagtaca agaggaagtg tatttgcatc agttgatact agaaaggtta
      781 gtagaaagga agaattcaga atttggattt cagttactct ttgtaaacgt tatgcttaat
      841 attatgaagt aactataata atcagcccat tgttttcatg cctagaaggt agctacttta
      901 ttatttcatt aacattttat gtatatacac atttttggtt ttaaaggtca ggaattaact
      961 tttgagattt atacccaaac agtctccttt cccagatttt aaattttaat ttggaatctt
     1021 ctaaacttct aattgaatta tttaatttct gaattctgac aacattaata taaaatacag
     1081 gacctattag aattacaagt cttttctttt tttttttttt ttttttttga gatagtttgt
     1141 ggctctcttg tcgaggctgg agtgcagtgg catgatctca gctcgctgca acctctgcct
     1201 cccaggttca agcaattctt gtgcctcaag tgtatctggg attacaggca tgtaccacca
     1261 tcctcagctg tttttttttt tgtactttta atagagatag gatttcacca tcttggctag
     1321 gctggtcttg aactcctggc ctcaagtgat ccacctgcct cagcctccca aattactggg
     1381 attataggca tgagccactg tgcccagcca caaggctacg agtctttttt ggtatgtcag
     1441 gtttttatag gcagcaccac ttttatacag aaacagtaag catgctaatt aaatatttat
     1501 gcttattcat tgagtgtagt ccattaagtt ttccccactt gccaaaacaa ttggcttaaa
     1561 aaatattttt cttgagacat tttttgcaat ttagacttcc tggtttgaca tgagcctttt
     1621 tttcaagtga atcagattta gtgagaatga agttctttgt atatttgggg aagacatggt
     1681 agataccttg agtccttgct ctcattctga gtgacgtaac tttaaagttg tcataaaatt
     1741 aaatgtttgt ttcttttctt ttcttttttc tttctttgtc ttctttcttt cttttttttt
     1801 ttttaaagaa ctaccagggc aagagcactt tgaacacagc aggattttct tcttcacaaa
     1861 tgtccaatcc tgtggacgac gaatcatggg atgctagcgc cacgaacttc tctctgttaa
     1921 agcaagcagg agacgtggaa gaaaaccccg gtcctgttag caagggcgag gaggtcatca
     1981 aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga
     2041 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     2101 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct
     2161 ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg
     2221 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc
     2281 aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact
     2341 tcccccccga cggccccgta atgcagaaaa agaccatggg ctgggaggcc tccaccgagc
     2401 gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg
     2461 acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac
     2521 tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca
     2581 ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg gggcatggca
     2641 ccggcagcac cggcagcggc agctccggca ccgcctcctc cgaggacaac aacatggccg
     2701 tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt
     2761 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     2821 aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttcatgt
     2881 acggctccaa ggcgtacgtg aagcaccccg ccgacatccc cgattacaag aagctgtcct
     2941 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg
     3001 tgacccagga ctcctccctg caggacggca cgctgatcta caaggtgaag atgcgcggca
     3061 ccaacttccc ccccgacggc cccgtaatgc agaaaaagac catgggctgg gaggcctcca
     3121 ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccaccag gccctgaagc
     3181 tgaaggacgg cggccactac ctggtggagt tcaagaccat ctacatggcc aagaagcccg
     3241 tgcaactgcc cggctactac tacgtggaca ccaagctgga catcacctcc cacaacgagg
     3301 actacaccat cgtggaacag tacgagcgct ccgagggccg ccaccacctg ttcctgtacg
     3361 gcatggacga gctgtacaag taagtcgact aaaactgaaa catccttcaa gtctgtagtt
     3421 ttgctgcaga gaagaaaata gttttcattt ttgagttttt aacagaagta taaaacctga
     3481 cattgtcata gctcctgtcc ttgtgttctc tctcctacac ttaaaaagaa atacatactg
     3541 actagttatg tgagatgcta aaagttacaa cattgcagtt actgatacaa atggtgttaa
     3601 ctagaaatat taaagcattc taaagtcttt cttatttctc atatattctt cagggggttt
     3661 agatatgttt aatgtttaaa tgccaagtct tactatagtg tttattgatc ttataaaaca
     3721 agtatctagg atatgataca cttttggtca aaaattactg ggtctcattt ttacttgagc
     3781 ctttaagaca gtagtgtgtc actataatgt gatagcatga tctttaagaa acagtagaag
     3841 acttaagcct ttcaaagtga ttttgatttt tagatcatca gatgtatgat gaaaatggtt
     3901 aaatgtttgt gacatgagcg ttgtactcaa tggcataaca atgtttgttt ttataatata
     3961 cactctttcc ttgaaataaa ggaagaaaca cttttccctt aagttttcca tcagcacttt
     4021 aaatttttct ttgttcagta tcctagcccc aactcctgta tttttgaaac tttgtttttt
     4081 tgtgtgtctt ttgtaaccaa tgtagtgatc aaagtttcac ataaattatt tatgtatagt
     4141 atacattact acacagatgt ggcaatgaaa atcaatctta agactcaaac ccatttatcc
     4201 acaaaaaaca gtgctgctta ctcttgccat aagttttgat ttatatctta agatcaatgg
     4261 aaagatccct gtgtttcttc attttccatt atatatctta aacttactat gctactatgc
     4321 ctagaatata cgaagttctt ttcttttttt tttttttttt tttgagacag agtcttgctc
     4381 ttatcaccca ggctggagta caatggtgca atcttggctc actgcagcct ctgcctcctg
     4441 tgttcaagca gttcttctgt cttagcctcc taattagctg ggactacaga tgtgcaccat
     4501 catgcctggc taagtattat atttttagta gagatgggct ttcaccacgt tggccaggct
     4561 ggtctcaaac tcttgacctc atgatccgcc cgccctggcc tctcaaagtg ctaagattac
     4621 aggcataagc cactgcgcct ggcctgaagt tcttaataaa ttgttaaatg aacgaaccca
     4681 aatgtttaac acctaatgag ttgtaaataa tggggagaca aaaatgattg gtattgttac
     4741 tgttcaccta agcaatccaa aaatgacatg tccctgatgt atatctggtt aaggaattta
     4801 gttaataaat gaggcatcac atatgaaaaa agagggcata tcaacagata gtgttaggat
     4861 aactggatag atggctacat cttgccatat tcaaaattaa cttcatgtga attatgaagt
     4921 cctttcccca ttgcttttgt ccagtttatc aaagatcagc tggttgtaga tgtgtggctt
     4981 tgcttctaag gtcaagggcg aattctgcag atatccatca cactggcggc cgctcgagca
     5041 tgcatctaga gggcccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt
     5101 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     5161 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     5221 gttgcgcagc ctatacgtac ggcagtttaa ggtttacacc tataaaagag agagccgtta
     5281 tcgtctgttt gtggatgtac agagtgatat tattgacacg ccggggcgac ggatggtgat
     5341 ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt
     5401 gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc
     5461 cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat
     5521 taacctgatg ttctggggaa tataaatgtc aggcatgaga ttatcaaaaa ggatcttcac
     5581 ctagatcctt ttcacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt
     5641 cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg
     5701 cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga
     5761 attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc
     5821 tttcttgccg ccaaggatct gatggcgcag gggatcaagc tctgatcaag agacaggatg
     5881 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     5941 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     6001 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     6061 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     6121 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     6181 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     6241 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     6301 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     6361 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     6421 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     6481 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     6541 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     6601 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     6661 ctatcgcctt cttgacgagt tcttctgaat tattaacgct tacaatttcc tgatgcggta
     6721 ttttctcctt acgcatctgt gcggtatttc acaccgcatc aggtggcact tttcggggaa
     6781 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6841 tgagacaata accctgataa atgcttcaat aatagcacgt gaggagggcc accatggcca
     6901 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     6961 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     7021 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     7081 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     7141 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     7201 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     7261 aggactgaca cgtgctaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7321 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7381 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7441 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7501 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     7561 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7621 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7681 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7741 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7801 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7861 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7921 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7981 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     8041 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     8101 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     8161 gcgaggaagc ggaag
//
