LOCUS       sequence-123710-        6564 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 6564)
  AUTHORS   Warren L, Manos PD, Ahfeldt T, Loh YH, Li H, Lau F, Ebina W, Mandal 
            PK, Smith ZD, Meissner A, Daley GQ, Brack AS, Collins JJ, Cowan C, 
            Schlaeger TM, Rossi DJ
  TITLE     Highly efficient reprogramming to pluripotency and directed 
            differentiation of human cells with synthetic modified mRNA.
  JOURNAL   Cell Stem Cell. 2010 Nov 5;7(5):618-30. doi: 
            10.1016/j.stem.2010.08.012. Epub 2010 Sep 30.
  PUBMED    20888316
REFERENCE   2  (bases 1 to 6564)
  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.stem.2010.08.012"; journalName: "Cell Stem Cell"; date: 
            "2010-11-5- 5"; volume: "7"; issue: "5"; pages: "618-30"
COMMENT     Sequence Label: pcDNA3.3_SOX2
FEATURES             Location/Qualifiers
     source          1..6564
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        40..419
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        420..623
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     574..594
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     620..644
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     752..771
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        752..770
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1940..1959)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1984..2032
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2234..2662
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2321..2340)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2531..2552
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2671..2691)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2676..3005
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2856..2991
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2918..2937
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3072..3866
                     /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(3126..3145)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3736..3755
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4042..4163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4079..4098)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4133..4152
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4212..4228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4212..4228)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4225..4247)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4236..4252
                     /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(4260..4266,4267..4284,4285..4290))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4305..4326
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4443..4460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4614..5202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4694..4713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5373..6164,6165..6233))
                     /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     5996..6015
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6234..6338)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6413..6432)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
       61 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      121 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      181 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      241 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      301 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      361 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      421 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      481 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      541 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      601 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
      661 cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg actctagagg
      721 atcgaaccct tttggaccct cgtacagaag ctaatacgac tcactatagg gaaataagag
      781 agaaaagaag agtaagaaga aatataagag ccaccatgta caacatgatg gagacggagc
      841 tgaagccgcc gggcccgcag caaacttcgg ggggcggcgg cggcaactcc accgcggcgg
      901 cggccggcgg caaccagaaa aacagcccgg accgcgtcaa gcggcccatg aatgccttca
      961 tggtgtggtc ccgcgggcag cggcgcaaga tggcccaaga gaaccccaag atgcacaact
     1021 cggagatcag caagcgcctg ggcgccgagt ggaaactttt gtcggagacg gagaagcggc
     1081 cgttcatcga cgaggctaag cggctgcgag cgctgcacat gaaggagcac ccggattata
     1141 aataccggcc ccggcggaaa accaagacgc tcatgaagaa ggataagtac acgctgcccg
     1201 gcgggctgct ggcccccggc ggcaatagca tggcgagcgg ggtcggggtg ggcgccggcc
     1261 tgggcgcggg cgtgaaccag cgcatggaca gttacgcgca catgaacggc tggagcaacg
     1321 gcagctacag catgatgcag gaccagctgg gctacccgca gcacccgggc ctcaatgcgc
     1381 acggcgcagc gcagatgcag cccatgcacc gctacgacgt gagcgccctg cagtacaact
     1441 ccatgaccag ctcgcagacc tacatgaacg gctcgcccac ctacagcatg tcctactcgc
     1501 agcagggcac ccctggcatg gctcttggct ccatgggttc ggtggtcaag tccgaggcca
     1561 gctccagccc ccctgtggtt acctcttcct cccactccag ggcgccctgc caggccgggg
     1621 acctccggga catgatcagc atgtatctcc ccggcgccga ggtgccggaa cccgccgccc
     1681 ccagcagact tcacatgtcc cagcactacc agagcggccc ggtgcccggc acggccatta
     1741 acggcacact gcccctctca cacatgtgag ctgccttctg cggggcttgc cttctggcca
     1801 tgcccttctt ctctcccttg cacctgtacc tcttggtctt tgaataaagc ctgagtagga
     1861 agtgagggtc tagaactagt gtcgacgcaa gggttcgatc cctaccggtt agtaatgagt
     1921 ttaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     1981 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     2041 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     2101 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     2161 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     2221 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2281 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2341 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2401 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2461 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2521 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     2581 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2641 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2701 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2761 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2821 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     2881 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     2941 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3001 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3061 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3121 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3181 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3241 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3301 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3361 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3421 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3481 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3541 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3601 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3661 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3721 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3781 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3841 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     3901 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3961 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4021 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4081 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4141 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4201 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4261 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4321 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4381 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4441 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4501 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4561 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4621 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4681 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4741 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4801 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4861 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4921 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4981 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5041 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5101 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5161 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5221 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5281 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5341 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5401 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5461 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5521 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5581 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5641 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5701 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5761 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5821 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5881 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5941 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6001 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6061 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6121 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6181 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6241 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6301 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6361 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     6421 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     6481 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     6541 agaatctgct tagggttagg cgtt
//
