LOCUS       sequence-375027-        6217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Polycistronic human LIN28 and NANOG were cloned into the pMXs 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6217)
  AUTHORS   Wang L, Su Y, Huang C, Yin Y, Chu A, Knupp A, Tang Y
  TITLE     NANOG and LIN28 dramatically improve human cell reprogramming by 
            modulating LIN41 and canonical WNT activities.
  JOURNAL   Biol Open. 2019 Dec 5;8(12). pii: 8/12/bio047225. doi: 
            10.1242/bio.047225.
  PUBMED    31806618
REFERENCE   2  (bases 1 to 6217)
  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: "Biol 
            Open."; date: "2019-12-5"; volume: "8(12). pii: 8/12/bio047225. 
            doi"; pages: " 10.1242/bio.047225"
COMMENT     Sequence Label: pMXs-LN2
FEATURES             Location/Qualifiers
     source          1..6217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    601..958
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1017..1433
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1363..1385
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1401..1417
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1443..1817
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1755..1771
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             2482..2538
                     /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"
     LTR             3645..4114
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4244..4261)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4415..5003)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4495..4514)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5167..5958,5959..6027))
                     /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     5790..5809
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6028..6132)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(6162..6217,1..538)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 aggcatggaa aaatacataa ctgagaatag aaaagttcag atcaaggtca ggaacagatg
       61 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
      121 ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
      181 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt
      241 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
      301 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
      361 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgatt gactgagtcg
      421 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
      481 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg
      541 ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggt
      601 aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga ctgattttat
      661 gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac
      721 tgacgagttc ggaacacccg gccgcaaccc tgggagacgt cccagggact tcgggggccg
      781 tttttgtggc ccgacctgag tccaaaaatc ccgatcgttt tggactcttt ggtgcacccc
      841 cctaatagga gggatatgtg gttctggtag gagacgagaa cctaaaacag ttcccgcctc
      901 cgtctgaatt tttgctttcg gtttgggacc gaagccgcgc cgcgcgtctt gtctgctgca
      961 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
     1021 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     1081 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     1141 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     1201 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     1261 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     1321 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     1381 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gcccccatat
     1441 ggccatatga gatcttatat ggggcacccc cgccccttgt aaacttccct gaccctgaca
     1501 tgacaagagt tactaacagc ccctctctcc aagctcactt acaggctctc tacttagtcc
     1561 agcacgaagt ctggagacct ctggcggcag cctaccaaga acaactggac cgaccggtgg
     1621 tacctcaccc ttaccgagtc ggcgacacag tgtgggtccg ccgacaccag actaagaacc
     1681 tagaacctcg ctggaaagga ccttacacag tcctgctgac cacccccacc gccctcaaag
     1741 tagacggcat cgcagcttgg atacacgccg cccacgtgaa ggctgccgac cccgggggtg
     1801 gaccatcctc tagactgccg gatctagcta gttaattaag gatccatggg ctccgtgtcc
     1861 aaccagcagt ttgcaggtgg ctgcgccaag gcggcagaag aggcgcccga ggaggcgccg
     1921 gaggacgcgg cccgggcggc ggacgagcct cagctgctgc acggtgcggg catctgtaag
     1981 tggttcaacg tgcgcatggg gttcggcttc ctgtccatga ccgcccgcgc cggggtcgcg
     2041 ctcgaccccc cagtggatgt ctttgtgcac cagagtaagc tgcacatgga agggttccgg
     2101 agcttgaagg agggtgaggc agtggagttc acctttaaga agtcagccaa gggtctggaa
     2161 tccatccgtg tcaccggacc tggtggagta ttctgtattg ggagtgagag gcggccaaaa
     2221 ggaaagagca tgcagaagcg cagatcaaaa ggagacaggt gctacaactg tggaggtcta
     2281 gatcatcatg ccaaggaatg caagctgcca ccccagccca agaagtgcca cttctgccag
     2341 agcatcagcc atatggtagc ctcatgtccg ctgaaggccc agcagggccc tagtgcacag
     2401 ggaaagccaa cctactttcg agaggaagaa gaagaaatcc acagccctac cctgctcccg
     2461 gaggcacaga atggatctgg agcaacaaac ttctcactac tcaaacaagc aggtgacgtg
     2521 gaggagaatc ccggccctat gagtgtggat ccagcttgtc cccaaagctt gccttgcttt
     2581 gaagcatccg actgtaaaga atcttcacct atgcctgtga tttgtgggcc tgaagaaaac
     2641 tatccatcct tgcaaatgtc ttctgctgag atgcctcaca cggagactgt ctctcctctt
     2701 ccctcctcca tggatctgct tattcaggac agccctgatt cttccaccag tcccaaaggc
     2761 aaacaaccca cttctgcaga gaatagtgtc gcaaaaaagg aagacaaggt cccagtcaag
     2821 aaacagaaga ccagaactgt gttctcttcc acccagctgt gtgtactcaa tgatagattt
     2881 cagagacaga aatacctcag tctccagcag atgcaagaac tctccaacat cctgaacctc
     2941 agctacaaac aggtgaagac ctggttccag aaccagagaa tgaaatctaa gaggtggcag
     3001 aaaaacaact ggccgaagaa tagcaatggt gtgacgcaga aggcctcagc acctacctac
     3061 cccagcctct actcttccta ccaccaggga tgcctggtga acccgactgg gaaccttcca
     3121 atgtggagca accagacctg gaacaattca acctggagca accagaccca gaacatccag
     3181 tcctggagca accactcctg gaacactcag acctggtgca cccaatcctg gaacaatcag
     3241 gcctggaaca gtcccttcta taactgtgga gaggaatctc tgcagtcctg catgcagttc
     3301 cagccaaatt ctcctgccag tgacttggag gctgctttgg aagctgctgg ggaaggcctt
     3361 aatgtaatac agcagaccac taggtatttt agtactccac aaaccatgga tttattccta
     3421 aactactcca tgaacatgca acctgaagac gtgtgagcgg ccgccagcac agtggtcgac
     3481 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     3541 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact
     3601 gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa
     3661 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc
     3721 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     3781 aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt
     3841 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     3901 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     3961 cctcactcgg sgcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
     4021 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     4081 gagtgattga ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt
     4141 ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga atcggccaac
     4201 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4261 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4321 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4381 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4441 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4501 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4561 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4621 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4681 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4741 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4801 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4861 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4921 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4981 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5041 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5101 cctagatcct tttgcggccg gccgcaaatc aatctaaagt atatatgagt aaacttggtc
     5161 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5221 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5281 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5341 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5401 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5461 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5521 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5581 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5641 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5701 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5761 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5821 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5881 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5941 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6001 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6061 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6121 aggggttccg cgcacatttc cccgaaaagt gccacctgca taatgaaaga ccccacctgt
     6181 aggtttggca agctagctta agtaacgcca ttttgca
//
