LOCUS       sequence-10636-e        8398 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 8398)
  AUTHORS   Papapetrou EP, Tomishima MJ, Chambers SM, Mica Y, Reed E, Menon J, 
            Tabar V, Mo Q, Studer L, Sadelain M
  TITLE     Stoichiometric and temporal requirements of Oct4, Sox2, Klf4, and 
            c-Myc expression for efficient human iPSC induction and 
            differentiation.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Aug 4. 106(31):12759-64.
  PUBMED    19549847
REFERENCE   2  (bases 1 to 8398)
  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: "Proc Natl 
            Acad Sci U S A. 2009 Aug 4. 106(31):12759-64."
COMMENT     Sequence Label: pLM-mCitrine-Sox2
FEATURES             Location/Qualifiers
     source          1..8398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    2..590
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(55..75)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(473..484)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             743..923
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    995..1116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1032..1051)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1086..1105
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1135..1270
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1302..1321)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1303..1321)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1331..1348)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1331..1347)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1340..1362)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1492..1947
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1579..1598)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1789..1810
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1976..2080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2081..2149,2150..2941)
                     /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     complement(2299..2318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3112..3700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3601..3620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3854..3871
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3988..4009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4024..4029,4030..4047,4048..4054)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4062..4078
                     /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     4067..4089
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4086..4102
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4086..4102
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4121..4141
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4123..4141
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             4397..4577
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4621..4746
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5239..5472
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5657..5701
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5968..6085
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6132..6642
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     6561..6579
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(6657..6659,6660..6662,6663..7373)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6702..6723)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6963..6982)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7310..7331
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7383..7439
                     /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"
ORIGIN
        1 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
       61 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
      121 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
      181 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
      241 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
      301 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
      361 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
      421 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
      481 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
      541 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattcg
      601 agctcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
      661 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tatcctgcta
      721 gctccggatc cggacctgta ctgggtctct ctggttagac cagatctgag cctgggagct
      781 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
      841 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
      901 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
      961 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     1021 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     1081 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     1141 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     1201 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     1261 tttggaggcc taggcttttg cgtcgagacg tacccaattc gccctatagt gagtcgtatt
     1321 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     1381 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     1441 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc gacgcgccct
     1501 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     1561 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     1621 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     1681 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     1741 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     1801 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     1861 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     1921 ttaacaaaat attaacgttt acaatttccc aggtggcact tttcggggaa atgtgcgcgg
     1981 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2041 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2101 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2161 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2221 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2281 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     2341 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     2401 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     2461 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     2521 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     2581 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     2641 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     2701 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     2761 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     2821 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     2881 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     2941 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3001 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3061 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3121 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3181 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3241 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3301 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3361 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3421 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3481 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3541 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3601 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3661 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3721 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3781 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3841 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     3901 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     3961 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     4021 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     4081 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
     4141 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc
     4201 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc
     4261 cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg
     4321 acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta
     4381 gctcgataca ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     4441 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     4501 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     4561 gtggaaaatc tctagcagtg gcgcccgaac agggacctga aagcgaaagg gaaaccagag
     4621 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
     4681 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
     4741 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
     4801 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
     4861 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
     4921 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
     4981 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
     5041 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag
     5101 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
     5161 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
     5221 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
     5281 tatgggcgca gcctcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
     5341 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
     5401 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
     5461 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
     5521 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
     5581 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     5641 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     5701 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     5761 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     5821 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag
     5881 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
     5941 gaacggatct cgacggtatc ggttaacttt taaaagaaaa ggggggattg gggggtacag
     6001 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
     6061 acaaattaca aaaattcaaa attttatcga tcacgagact agcctcgaga agcttgatat
     6121 cgaattccca cggggttggg gttgcgcctt ttccaaggca gccctgggtt tgcgcaggga
     6181 cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca
     6241 ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt gggccccccg
     6301 gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg
     6361 acgtgacaaa cggaagccgc acgtctcact agtaccctcg cagacggaca gcgccaggga
     6421 gcaatggcag cgcgccgacc gcgatgggct gtggccaata gcggctgctc agcggggcgc
     6481 gccgagagca gcggccggga aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg
     6541 gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc ctccggagcg cacgtcggca
     6601 gtcggctccc tcgttgaccg aatcaccgac ctctctcccc agggggatcc accggtatgg
     6661 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     6721 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     6781 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     6841 tgaccacctt cggctacggc ctgatgtgct tcgcccgcta ccccgaccac atgaagcagc
     6901 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     6961 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     7021 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     7081 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     7141 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     7201 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     7261 tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     7321 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggatctg
     7381 gagcaacaaa cttctcacta ctcaaacaag caggtgacgt ggaggagaat cccggcccta
     7441 tgtacaacat gatggagacg gagctgaagc cgccgggccc gcagcaaact tcggggggcg
     7501 gcggcggcaa ctccaccgcg gcggcggccg gcggcaacca gaaaaacagc ccggaccgcg
     7561 tcaagcggcc catgaatgcc ttcatggtgt ggtcccgcgg gcagcggcgc aagatggccc
     7621 aggagaaccc caagatgcac aactcggaga tcagcaagcg cctgggcgcc gagtggaaac
     7681 ttttgtcgga gacggagaag cggccgttca tcgacgaggc taagcggctg cgagcgctgc
     7741 acatgaagga gcacccggat tataaatacc ggccccggcg gaaaaccaag acgctcatga
     7801 agaaggataa gtacacgctg cccggcgggc tgctggcccc cggcggcaat agcatggcga
     7861 gcggggtcgg ggtgggcgcc ggcctgggcg cgggcgtgaa ccagcgcatg gacagttacg
     7921 cgcacatgaa cggctggagc aacggcagct acagcatgat gcaggaccag ctgggctacc
     7981 cgcagcaccc gggcctcaat gcgcacggcg cagcgcagat gcagcccatg caccgctacg
     8041 acgtgagcgc cctgcagtac aactccatga ccagctcgca gacctacatg aacggctcgc
     8101 ccacctacag catgtcctac tcgcagcagg gcacccctgg catggctctt ggctccatgg
     8161 gttcggtggt caagtccgag gccagctcca gcccccctgt ggttacctct tcctcccact
     8221 ccagggcgcc ctgccaggcc ggggacctcc gggacatgat cagcatgtat ctccccggcg
     8281 ccgaggtgcc ggaacccgcc gcccccagca gacttcacat gtcccagcac taccagagcg
     8341 gcccggtgcc cggcacggcc attaacggca cactgcccct ctcacacatg tgagtcga
//
