LOCUS       sequence-169366-        6240 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 6240)
  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 6240)
  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_LIN28A
FEATURES             Location/Qualifiers
     source          1..6240
                     /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"
     polyA_signal    270..391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(281..300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     335..354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(567..1361)
                     /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(678..697)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1288..1307
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1428..1757)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1442..1577
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1496..1515)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1742..1762
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1771..2199)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1881..1902)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2093..2112
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2401..2449
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2474..2493
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     primer_bind     2565..2584
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2565..2583
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2623..2632
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3465..3489)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(3486..3689)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3515..3535)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3690..4069
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4241..4260
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4335..4439
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4440..4508,4509..5300)
                     /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(4658..4677)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5471..6059
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5960..5979
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6213..6230
                     /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 gccaagctct
      241 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
      301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      361 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
      421 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
      481 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
      541 tcatttcgcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
      601 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
      661 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
      721 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
      781 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
      841 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
      901 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
      961 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     1021 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     1081 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     1141 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
     1201 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     1261 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     1321 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     1381 gtctcttgat cagatccgaa aatggatata caagctcccg ggagcttttt gcaaaagcct
     1441 aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggcaga ggcggcctcg
     1501 gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc
     1561 ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga
     1621 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct
     1681 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     1741 caccctaact gacacacatt ccacagaatt aattcgcgtt aaatttttgt taaatcagct
     1801 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     1861 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     1921 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     1981 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     2041 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     2101 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     2161 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtg gggatacccc ctagagcccc
     2221 agctgcgcag atctgctatg gcagggcctg ccgccccgac gttggctgcg agccctgggc
     2281 cttcacccga acttgggggg tggggtgggg aaaaggaaga aacgcgggcg tattggcccc
     2341 aatggggtct cggtggggta tcgacagagt gccagccctg ggaccgaacc ccgcgtttat
     2401 gaacaaacga cccaacaccc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg
     2461 gttccttccg gtattgtctc cttccgtgtt tcagttagcc tcccccgttt aaactcatta
     2521 ctaaccggta gggatcgaac ccttttggac cctcgtacag aagctaatac gactcactat
     2581 agggaaataa gagagaaaag aagagtaaga agaaatataa gagccaccat gggctccgtg
     2641 tccaaccagc agtttgcagg tggctgcgcc aaggcggcag aagaggcgcc cgaggaggcg
     2701 ccggaggacg cggcccgggc ggcggacgag cctcagctgc tgcacggtgc gggcatctgt
     2761 aagtggttca acgtgcgcat ggggttcggc ttcctgtcca tgaccgcccg cgccggggtc
     2821 gcgctcgacc ccccagtgga tgtctttgtg caccagagta agctgcacat ggaagggttc
     2881 cggagcttga aggagggtga ggcagtggag ttcaccttta agaagtcagc caagggtctg
     2941 gaatccatcc gtgtcaccgg acctggtgga gtattctgta ttgggagtga gaggcggcca
     3001 aaaggaaaga gcatgcagaa gcgcagatca aaaggagaca ggtgctacaa ctgtggaggt
     3061 ctagatcatc atgccaagga atgcaagctg ccaccccagc ccaagaagtg ccacttctgc
     3121 cagagcatca gccatatggt agcctcatgt ccgctgaagg cccagcaggg ccctagtgca
     3181 cagggaaagc caacctactt tcgagaggaa gaagaagaaa tccacagccc taccctgctc
     3241 ccggaggcac agaattgagc tgccttctgc ggggcttgcc ttctggccat gcccttcttc
     3301 tctcccttgc acctgtacct cttggtcttt gaataaagcc tgagtaggaa gtgagggtct
     3361 agaactagtg tcgacgcaag ggttcgatcc tctagagtcc ggaggctgga tcggtcccgg
     3421 tgtcttctat ggaggtcaaa acagcgtgga tggcgtctcc aggcgatctg acggttcact
     3481 aaacgagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
     3541 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
     3601 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
     3661 ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg
     3721 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     3781 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     3841 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     3901 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     3961 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     4021 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca acgcgtatat
     4081 ctggcccgta catcgcgaag cagcgcaaaa cgcctaaccc taagcagatt cttcatgcaa
     4141 ttgtcggtca agccttgcct tgttgtagct taaattttgc tcgcgcacta ctcagcgacc
     4201 tccaacacac aagcagggag cagatactgg cttaactatg cggcatcaga gcagattgta
     4261 ctgagagtgc accatagggg atcgggagat ctcccgatcc gtcgacgtca ggtggcactt
     4321 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4381 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4441 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4501 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4561 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4621 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4681 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4741 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4801 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4861 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4921 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4981 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5041 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5101 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5161 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5221 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5281 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5341 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5401 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5461 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5521 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5581 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5641 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5701 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5761 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5821 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5881 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5941 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6001 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6061 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6121 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6181 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
