LOCUS       Exported                8142 bp ds-DNA     circular SYN 20-OCT-2022
DEFINITION  Polycistronic human Glis1, LIN28 and NANOG were cloned into the pMXs
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    pMXs-GNL
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8142)
  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 8142)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 20, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8142
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..594
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    657..1014
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1073..1489
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1419..1441
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1457..1473
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1499..1873
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1811..1827
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             2538..2591
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3516..3572
                     /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             5626..6095
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6225..6242)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6396..6984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6476..6495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7148..8008)
                     /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     7771..7790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8009..8113)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaaat acataactga gaatagaaaa gttcagatca aggtcaggaa cagatggaac
      121 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
      181 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
      241 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
      301 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
      361 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
      421 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg
      481 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
      541 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
      601 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
      661 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc
      721 ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
      781 gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt
      841 tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg caccccccta
      901 ataggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
      961 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat
     1021 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac
     1081 tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca
     1141 caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc
     1201 aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt
     1261 taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt
     1321 gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa
     1381 gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac
     1441 cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccc ccatatggcc
     1501 atatgagatc ttatatgggg cacccccgcc ccttgtaaac ttccctgacc ctgacatgac
     1561 aagagttact aacagcccct ctctccaagc tcacttacag gctctctact tagtccagca
     1621 cgaagtctgg agacctctgg cggcagccta ccaagaacaa ctggaccgac cggtggtacc
     1681 tcacccttac cgagtcggcg acacagtgtg ggtccgccga caccagacta agaacctaga
     1741 acctcgctgg aaaggacctt acacagtcct gctgaccacc cccaccgccc tcaaagtaga
     1801 cggcatcgca gcttggatac acgccgccca cgtgaaggct gccgaccccg ggggtggacc
     1861 atcctctaga ctgccggatc tagctagtta attaaggatc catgggctcc gtgtccaacc
     1921 agcagtttgc aggtggctgc gccaaggcgg cagaagaggc gcccgaggag gcgccggagg
     1981 acgcggcccg ggcggcggac gagcctcagc tgctgcacgg tgcgggcatc tgtaagtggt
     2041 tcaacgtgcg catggggttc ggcttcctgt ccatgaccgc ccgcgccggg gtcgcgctcg
     2101 accccccagt ggatgtcttt gtgcaccaga gtaagctgca catggaaggg ttccggagct
     2161 tgaaggaggg tgaggcagtg gagttcacct ttaagaagtc agccaagggt ctggaatcca
     2221 tccgtgtcac cggacctggt ggagtattct gtattgggag tgagaggcgg ccaaaaggaa
     2281 agagcatgca gaagcgcaga tcaaaaggag acaggtgcta caactgtgga ggtctagatc
     2341 atcatgccaa ggaatgcaag ctgccacccc agcccaagaa gtgccacttc tgccagagca
     2401 tcagccatat ggtagcctca tgtccgctga aggcccagca gggccctagt gcacagggaa
     2461 agccaaccta ctttcgagag gaagaagaag aaatccacag ccctaccctg ctcccggagg
     2521 cacagaatgg atctggagag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga
     2581 atcccggccc catgagtgtg gatccagctt gtccccaaag cttgccttgc tttgaagcat
     2641 ccgactgtaa agaatcttca cctatgcctg tgatttgtgg gcctgaagaa aactatccat
     2701 ccttgcaaat gtcttctgct gagatgcctc acacggagac tgtctctcct cttccctcct
     2761 ccatggatct gcttattcag gacagccctg attcttccac cagtcccaaa ggcaaacaac
     2821 ccacttctgc agagaatagt gtcgcaaaaa aggaagacaa ggtcccagtc aagaaacaga
     2881 agaccagaac tgtgttctct tccacccagc tgtgtgtact caatgataga tttcagagac
     2941 agaaatacct cagtctccag cagatgcaag aactctccaa catcctgaac ctcagctaca
     3001 aacaggtgaa gacctggttc cagaaccaga gaatgaaatc taagaggtgg cagaaaaaca
     3061 actggccgaa gaatagcaat ggtgtgacgc agaaggcctc agcacctacc taccccagcc
     3121 tctactcttc ctaccaccag ggatgcctgg tgaacccgac tgggaacctt ccaatgtgga
     3181 gcaaccagac ctggaacaat tcaacctgga gcaaccagac ccagaacatc cagtcctgga
     3241 gcaaccactc ctggaacact cagacctggt gcacccaatc ctggaacaat caggcctgga
     3301 acagtccctt ctataactgt ggagaggaat ctctgcagtc ctgcatgcag ttccagccaa
     3361 attctcctgc cagtgacttg gaggctgctt tggaagctgc tggggaaggc cttaatgtaa
     3421 tacagcagac cactaggtat tttagtactc cacaaaccat ggatttattc ctaaactact
     3481 ccatgaacat gcaacctgaa gacgtgggat ctggagcaac aaacttctca ctactcaaac
     3541 aagcaggtga cgtggaggag aatcccggcc ctatggcaga ggcccgcaca tccctgtctg
     3601 cccactgtcg gggcccgctg gccactggcc tgcacccaga cctggacctc ccgggccgaa
     3661 gcctcgccac ccctgcgcct tcctgctacc ttctgggcag cgaacccagc tctggcctgg
     3721 gcctccagcc cgagacccac ctccccgagg gcagcctgaa gcggtgctgt gtcttgggcc
     3781 taccccccac ctccccagcc tcctcctcac cctgtgcctc ctccgacgtc acctccatca
     3841 tccgctcctc ccagacgtct ctggtcacct gtgtaaatgg actccggagc ccccctctgg
     3901 cgggagatct ggggggccct tccaagcggg cccggcctgg ccctgcatcg acggacagcc
     3961 atgagggcag cttgcaactt gaagcctgcc ggaaggcgag cttcctgaag caggaacccg
     4021 cggatgagtt ttcagagctc tttgggcctc accagcaggg cctgccgccc ccctatcccc
     4081 tgtctcagtt gccgcctggc ccaagccttg gaggcctggg gctgggcctg ggaggcaggg
     4141 tggtggccgg gcggcaggcg tgccgctggg tggactgctg tgcagcctat gagcagcagg
     4201 aggagctggt gcggcacatc gagaagagcc acaccgacca gcgcaagggc gaggacttca
     4261 cctgcttctg ggctggctgc gtgcgccgct acaagccctt caacgcccgc tacaagctgc
     4321 tcatccacat gcgagtgcac tcgggcgaga agcccaacaa gtgcatgttt gaaggctgca
     4381 gcaaggcctt ctcacggctg gagaacctca agatccacct gaggagccac acgggcgaga
     4441 agccgtacct gtgccagcac ccgggttgcc agaaggcctt cagcaactcc agcgaccgcg
     4501 ccaagcacca gcgcacccac ctagacacga agccgtacgc ctgtcagatc cctggctgct
     4561 ccaagcgcta cacagacccc agctccctcc gcaagcacgt caaggcccat tcagccaaag
     4621 agcagcaggt gcgtaagaag ctgcatgcgg gccctgacac cgaggccgac gtcctgaccg
     4681 agtgtctggt cctgcagcag ctccacacgt ccacacagct ggctgccagc gacggcaagg
     4741 gtggctgtgg cctgggccag gagctgctcc caggtgtgta tcctggctcc atcacccccc
     4801 ataacggact tgcatcgggc ctcctgcccc cagcgcacga cgtaccttcc aggcaccacc
     4861 cgctggatgc caccaccagt tcccaccacc atctgtcccc tctgcccatg gctgagagca
     4921 cccgggatgg gttggggccc ggcctcctct caccaatagt cagccccctg aaggggctgg
     4981 ggccaccgcc gctgccccca tcctctcaga gccattctcc ggggggccag cccttcccca
     5041 cactccccag caagccgtcc tacccnaccc ttccagagcc ctccnacccc cgcctctgcc
     5101 cagcccacaa ggttaccagg gcagtttcca ctccatccag agttgcttcc cctatggcga
     5161 ctgctaccgg atggctgaac cagcagccgg tggggacgga ctggtcgggg agacccacgg
     5221 tttcaacccc ctgcggccca atggctacca cagcctcagc acgcccttgc ctgccacagg
     5281 ctatgaggcc ctggctgagg cctcatgccc cacagcgctg ccacagcagc catctgaaga
     5341 tgtggtgtcc agcggccccg aggactgtgg cttcttcccc aatggagcct ttgaccactg
     5401 cctgggccac atcccctcca tctacacaga cacctgagcg gccgccagca cagtggtcga
     5461 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     5521 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
     5581 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
     5641 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
     5701 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     5761 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
     5821 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     5881 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     5941 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     6001 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     6061 tgagtgattg actacccgtc agcgggggtc tttcacatgc agcatgtatc aaaattaatt
     6121 tggttttttt tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa
     6181 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6241 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6301 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6361 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6421 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6481 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6541 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6601 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6661 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6721 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6781 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6841 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6901 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6961 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7021 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7081 acctagatcc ttttgcggcc ggccgcaaat caatctaaag tatatatgag taaacttggt
     7141 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7201 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7261 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7321 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7381 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7441 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7501 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7561 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7621 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7681 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7741 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7801 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7861 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7921 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7981 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8041 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8101 taggggttcc gcgcacattt ccccgaaaag tgccacctgc at
//
