LOCUS       sequence-123707-        7023 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 7023)
  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 7023)
  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_KLF4
FEATURES             Location/Qualifiers
     source          1..7023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        133..512
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        513..716
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     667..687
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     713..737
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     845..864
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        845..863
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      903..912
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2492..2511)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2536..2584
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2786..3214
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2873..2892)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3083..3104
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3223..3243)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3228..3557
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3408..3543
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3470..3489
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3624..4418
                     /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(3678..3697)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4288..4307
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4594..4715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4631..4650)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4685..4704
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4764..4780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4764..4780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4777..4799)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4788..4804
                     /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(4812..4818,4819..4836,4837..4842))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4857..4878
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4995..5012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5166..5754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5246..5265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5925..6716,6717..6785))
                     /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     6548..6567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6786..6890)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6965..6984)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
       61 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
      121 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      181 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      241 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      301 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      361 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      421 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      481 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      541 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      601 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      661 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
      721 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
      781 accgggaccg atccagcctc cggactctag aggatcgaac ccttttggac cctcgtacag
      841 aagctaatac gactcactat agggaaataa gagagaaaag aagagtaaga agaaatataa
      901 gagccaccat ggctgtcagc gacgcgctgc tcccatcttt ctccacgttc gcgtctggcc
      961 cggcgggaag ggagaagaca ctgcgtcaag caggtgcccc gaataaccgc tggcgggagg
     1021 agctctccca catgaagcga cttcccccag tgcttcccgg ccgcccctat gacctggcgg
     1081 cggcgaccgt ggccacagac ctggagagcg gcggagccgg tgcggcttgc ggcggtagca
     1141 acctggcgcc cctacctcgg agagagaccg aggagttcaa cgatctcctg gacctggact
     1201 ttattctctc caattcgctg acccatcctc cggagtcagt ggccgccacc gtgtcctcgt
     1261 cagcgtcagc ctcctcttcg tcgtcgccgt cgagcagcgg ccctgccagc gcgccctcca
     1321 cctgcagctt cacctatccg atccgggccg ggaacgaccc gggcgtggcg ccgggcggca
     1381 cgggcggagg cctcctctat ggcagggagt ccgctccccc tccgacggct cccttcaacc
     1441 tggcggacat caacgacgtg agcccctcgg gcggcttcgt ggccgagctc ctgcggccag
     1501 aattggaccc ggtgtacatt ccgccgcagc agccgcagcc gccaggtggc gggctgatgg
     1561 gcaagttcgt gctgaaggcg tcgctgagcg cccctggcag cgagtacggc agcccgtcgg
     1621 tcatcagcgt cagcaaaggc agccctgacg gcagccaccc ggtggtggtg gcgccctaca
     1681 acggcgggcc gccgcgcacg tgccccaaga tcaagcagga ggcggtctct tcgtgcaccc
     1741 acttgggcgc tggaccccct ctcagcaatg gccaccggcc ggctgcacac gacttccccc
     1801 tggggcggca gctccccagc aggactaccc cgaccctggg tcttgaggaa gtgctgagca
     1861 gcagggactg tcaccctgcc ctgccgcttc ctcccggctt ccatccccac ccggggccca
     1921 attacccatc cttcctgccc gatcagatgc agccgcaagt cccgccgctc cattaccaag
     1981 agctcatgcc acccggttcc tgcatgccag aggagcccaa gccaaagagg ggaagacgat
     2041 cgtggccccg gaaaaggacc gccacccaca cttgtgatta cgcgggctgc ggcaaaacct
     2101 acacaaagag ttcccatctc aaggcacacc tgcgaaccca cacaggtgag aaaccttacc
     2161 actgtgactg ggacggctgt ggatggaaat tcgcccgctc agatgaactg accaggcact
     2221 accgtaaaca cacggggcac cgcccgttcc agtgccaaaa atgcgaccga gcattttcca
     2281 ggtcggacca cctcgcctta cacatgaaga ggcattttta agctgccttc tgcggggctt
     2341 gccttctggc catgcccttc ttctctccct tgcacctgta cctcttggtc tttgaataaa
     2401 gcctgagtag gaagtgaggg tctagaacta gtgtcgacgc aagggttcga tccctaccgg
     2461 ttagtaatga gtttaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     2521 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
     2581 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     2641 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     2701 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc
     2761 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2821 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2881 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2941 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3001 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3061 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3121 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3181 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3241 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3301 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3361 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3421 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3481 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3541 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3601 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3661 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3721 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3781 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3841 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3901 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3961 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4021 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4081 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4141 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4201 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4261 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4321 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4381 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa
     4441 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4501 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4561 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4621 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4681 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4741 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4801 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4861 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4921 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4981 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5041 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5101 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5161 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5221 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5281 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5341 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5401 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5461 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5521 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5581 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5641 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5701 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5761 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5821 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5881 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5941 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6001 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6061 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6121 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6181 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6241 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6301 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6361 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6421 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6481 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6541 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6601 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6661 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6721 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6781 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6841 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6901 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     6961 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     7021 tgg
//
