LOCUS       Exported                7227 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Transient human NANOG expression.
ACCESSION   .
VERSION     .
KEYWORDS    PpyCAG-NANOG-IB
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7227)
  AUTHORS   Takashima Y, Guo G, Loos R, Nichols J, Ficz G, Krueger F, Oxley D, 
            Santos F, Clarke J, Mansfield W, Reik W, Bertone P, Smith A
  TITLE     Resetting Transcription Factor Control Circuitry toward Ground-State
            Pluripotency in Human.
  JOURNAL   Cell. 2014 Sep 11;158(6):1254-69. doi: 10.1016/j.cell.2014.08.029.
  PUBMED    25215486
REFERENCE   2  (bases 1 to 7227)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7227
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    1..25
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(949..973)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1107..1692
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1274..1291)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1501..1520
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(2105..2122)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2111..2335
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2382..2398)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2382..2398)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2395..2417)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2406..2422
                     /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(2430..2460)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2475..2496
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2555..2750
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2601..2736
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2663..2682
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2756..2890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2958..2975)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3129..3717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3209..3228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3888..4748)
                     /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     4511..4530
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5351..5485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        5499..5878
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5881..6156
                     /label=chicken beta-actin promoter
     intron          6157..7165
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7089..7111
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 acaagtttgt acaaaaaagc aggctcacca tgagtgtgga tccagcttgt ccccaaagct
       61 tgccttgctt tgaagcatcc gactgtaaag aatcttcacc tatgcctgtg atttgtgggc
      121 ctgaagaaaa ctatccatcc ttgcaaatgt cttctgctga gatgcctcac acggagactg
      181 tctctcctct tccctcctcc atggatctgc ttattcagga cagccctgat tcttccacca
      241 gtcccaaagg caaacaaccc acttctgcag agaatagtgt cgcaaaaaag gaagacaagg
      301 tcccagtcaa gaaacagaag accagaactg tgttctcttc cacccagctg tgtgtactca
      361 atgatagatt tcagagacag aaatacctca gcctccagca gatgcaagaa ctctccaaca
      421 tcctgaacct cagctacaaa caggtgaaga cctggttcca gaaccagaga atgaaatcta
      481 agaggtggca gaaaaacaac tggccgaaga atagcaatgg tgtgacgcag aaggcctcag
      541 cacctaccta ccccagcctc tactcttcct accaccaggg atgcctggtg aacccgactg
      601 ggaaccttcc aatgtggagc aaccagacct ggaacaattc aaccaggagc aaccagaccc
      661 agaacatcca gtcctggagc aaccactcct ggaacactca gacctggtgc acccaatcct
      721 ggaacaatca ggcctggaac agtcccttct ataactgtgg agaggaatct ctgcagtcct
      781 gcatgcagtt ccagccaaat tctcctgcca gtgacttgga ggctgctttg gaagctgctg
      841 gggaaggcct taatgtaata cagcagacca ctaggtattt tagtactcca caaaccatgg
      901 atttattcct aaactactcc atgaacatgc aacctgaaga cgtgtgagac ccagctttct
      961 tgtacaaagt ggtgattcga ggaattcact cctcaggtgc aggctgccta tcagaaggtg
     1021 gtggctggtg tggccaactg gctcacaaat accactgaga tcgacggtat cgagcggccg
     1081 ctcgataagc ttgatatcga attccgcccc tctccctccc ccccccctaa cgttactggc
     1141 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     1201 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     1261 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     1321 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     1381 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     1441 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     1501 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     1561 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     1621 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     1681 atggccacaa ccatgccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct
     1741 acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc
     1801 gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa
     1861 ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg
     1921 atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc
     1981 gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt
     2041 gggattcgtg aattgctgcc ctctggttat gtgtgggagg gttaactaga taagtaatga
     2101 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2161 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2221 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2281 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2341 gaggcggaaa gaacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg
     2401 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     2461 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2521 ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag
     2581 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2641 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     2701 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt
     2761 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     2821 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     2881 tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2941 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3001 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3061 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3121 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3181 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3241 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3301 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     3361 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3421 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3481 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3541 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     3601 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3661 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3721 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3781 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3841 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3901 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3961 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4021 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4081 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4141 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4201 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4261 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4321 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4381 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4441 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4501 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4561 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4621 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4681 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4741 atactcatac tcttcctttt tcaatgaccg ctttggaaaa acaaagactg tatttcctgg
     4801 aaattaatgt ttattcaata aactgtgtat tcagctatat cacatagtgg tgaggctgaa
     4861 atgaggcggg aagaggcggt tggggcttaa ttatatcaat ttgggtggcc ccacagcgcc
     4921 tccaaggcgc cagtcctgtt ttgacaagtt gcctctggaa gcctctacaa tgcctctctt
     4981 ctttttctcc agagtaagcg gaggccaggg gcccccggcc tctgcttaat actaaaaaaa
     5041 acagctgttg tcatagtaat gattgggtgg aaacattcca ggcctgggtg gagaggcttt
     5101 ttgcttcctc ttgcaaaacc acactgacat tccaggcctg ggtggagagg ctttttgctt
     5161 cctcttgcaa aaccacactg ccctctggag ggcagttgcc tagcaactaa ctaaaagagg
     5221 atgtcgcacg gccagctgcg gtcagttagt cacttcctgc ttaactgact tgacattttc
     5281 tattttaaga gtcgggagga aaattactgt gttggaggcc ctccgccatc ttctgaagct
     5341 gaatcgaatt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5401 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5461 caatgtatct tatcatgtct ggatctgata tcatcgtcga cattgattat tgactagtta
     5521 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     5581 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     5641 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     5701 ggactattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     5761 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     5821 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggg
     5881 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
     5941 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
     6001 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
     6061 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
     6121 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgtt gccttcgccc
     6181 cgtgccccgc tccgcgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
     6241 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
     6301 tgacggctcg tttcttttct gtggctgcgt gaaagcctta aagggctccg ggagggccct
     6361 ttgtgcgggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc
     6421 gtgcggcccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg
     6481 ctccgcgtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc gggggggctg
     6541 cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg
     6601 cgcggcggtc gggctgtaac ccccccctgc acccccctcc ccgagttgct gagcacggcc
     6661 cggcttcggg tgcggggctc cgtgcggggc gtggcgcggg gctcgccgtg ccgggcgggg
     6721 ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg
     6781 gggaggggcg cggcggcccc ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca
     6841 ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc aaatctggcg
     6901 gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcgggcg aagcggtgcg
     6961 gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc
     7021 ttctccatct ccagcctcgg ggctgccgca gggggacggc tgccttcggg ggggacgggg
     7081 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
     7141 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttgttg tgctgtctca
     7201 tcattttggc aaagaattcc tcgaatc
//
