LOCUS       sequence-147136-        7764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mNanog Homeodomain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7764)
  AUTHORS   Theunissen TW, Costa Y, Radzisheuskaya A, van Oosten AL, Lavial F, 
            Pain B, Castro LF, Silva JC
  TITLE     Reprogramming capacity of Nanog is functionally conserved in 
            vertebrates and resides in a unique homeodomain.
  JOURNAL   Development. 2011 Nov;138(22):4853-65. doi: 10.1242/dev.068775.
  PUBMED    22028025
REFERENCE   2  (bases 1 to 7764)
  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: "10.1242/dev"; 
            journalName: "Development"; date: "2011-11"; volume: "138"; issue: 
            "22"; pages: "4853-65"
COMMENT     Sequence Label: pPB-CAG-mNanogHD-pgk-hph
FEATURES             Location/Qualifiers
     source          1..7764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..658
                     /label=chicken beta-actin promoter
     intron          659..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1748..1772
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1995..2019)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2040..2056)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2064..2080
                     /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(2088..2094,2095..2112,2113..2118))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2133..2154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2214..2231)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2220..2444
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    2521..2554
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2777..2796)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2778..2796)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2806..2823)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2806..2822)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2815..2837)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2964..3419
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3051..3070)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3261..3282
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3445..3549
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3550..3618,3619..4410)
                     /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(3768..3787)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4581..5169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5070..5089
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5323..5340
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5457..5478
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5493..5498,5499..5516,5517..5523)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5531..5547
                     /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     5555..5571
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5590..5610
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5592..5610
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   5633..5667
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     CDS             complement(6150..7175)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(6150..7175)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(7194..7693)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     7635..7658
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    7716..7749
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      421 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      481 atgggggcgg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      661 agtcgctgcg ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg
      721 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      781 ctgtaattag cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct
      841 taaagggctc cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt
      901 gtgtgtgcgt ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg
      961 gcgcggcgcg gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt
     1021 gccccgcggt gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     1081 gggggtgagc agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct
     1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg
     1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc
     1321 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     1381 ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta
     1441 gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc
     1501 gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcctcgac ggtatcgata
     1741 agctatcaca agtttgtaca aaaaagcagg cttcaccatg aaggtccttg ccaggaagca
     1801 gaagatgcgg actgtgttct ctcaggccca gctgtgtgca ctcaaggaca ggtttcagaa
     1861 gcagaagtac ctcagcctcc agcagatgca agaactctcc tccattctga acctgagcta
     1921 taagcaggtt aagacctggt ttcaaaacca aaggatgaag tgcaagcggt ggcagaaaaa
     1981 ccagtggttg taagacccag ctttcttgta caaagtggtg atagcttggc gtaatcatgg
     2041 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2101 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2161 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat cagcctcgac
     2221 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2281 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2341 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2401 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2461 aaccagctgg ggctcgagat ccactagttc tagcctcgag gctagagcgg ccgcgtcgag
     2521 ataacttcgt ataatgtatg ctatacgaag ttatgttttg ttactttata gaagaaattt
     2581 tgagtttttg ttttttttta ataaataaat aaacataaat aaattgtttg ttgaatttat
     2641 tattagtatg taagtgtaaa tataataaaa cttaatatct attcaaatta ataaataaac
     2701 gtcaatttta cgcatgatta tctttaacgt acgtcacaat atgattatct ttctagggtt
     2761 aaagagctcc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     2821 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2881 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2941 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3001 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3061 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3121 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3181 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3241 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3301 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3361 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     3421 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3481 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3541 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3601 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3661 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3721 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3781 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3841 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3901 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3961 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4021 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4081 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4141 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4201 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4261 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4321 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4381 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4441 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4501 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4561 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4621 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4681 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     4741 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4801 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4861 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4921 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4981 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5041 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5101 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5161 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5221 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5281 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5341 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     5401 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     5461 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     5521 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     5581 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tgggtacctt aaccctagaa
     5641 agatagtctg cgtaaaattg acgcatgcat tcttgaaata ttgctctctc tttctaaata
     5701 gcgcgaatcc gtcgctgtgc atttaggaca tctcagtcgc cgcttggagc tcccgtgagg
     5761 cgtgcttgtc aatgcggtaa gtgtcactga ttttgaacta taacgaccgc gtgagtcaaa
     5821 atgacgcatg attatctttt acgtgacttt taagatttaa ctcatacgat aattatattg
     5881 ttatttcatg ttctacttac gtgataactt attatatata tattttcttg ttatagatat
     5941 ccaactatga aacattatca taaagcaata gtaaagagcc ttcagtaaag agcaggcatt
     6001 tatctaatcc caccccaccc ccacccccgt agctccaatc cttccattca aaatgtaggt
     6061 actctgttct cacccttctt aacaaagtat gacaggaaaa aacttccatg tggacatctt
     6121 tattgtttaa tagatcatca atttctcgac tattcctttg ccctcggacg agtgctgggg
     6181 cgtcggtttc cactatcggc gagtacttct acacagccat cggtccagac ggccgcgctt
     6241 ctgcgggcga tttgtgtacg cccgacagtc ccggctccgg atcggacgat tgcgtcgcat
     6301 cgaccctgcg cccaagctgc atcatcgaaa ttgccgtcaa ccaagctctg atagagttgg
     6361 tcaagaccaa tgcggagcat atacgcccgg agccgcggcg atcctgcaag ctccggatgc
     6421 ctccgctcga agtagcgcgt ctgctgctcc atacaagcca accacggcct ccagaagaag
     6481 atgttggcga cctcgtattg ggaatccccg aacatcgcct cgctccagtc aatgaccgct
     6541 gttatgcggc cattgtccgt caggacattg ttggagccga aatccgcgtg cacgaggtgc
     6601 cggacttcgg ggcagtcctc ggcccaaagc atcagctcat cgagagcctg cgcgacggac
     6661 gcactgacgg tgtcgtccat cacagtttgc cagtgataca catggggatc agcaatcgcg
     6721 catatgaaat cacgccatgt agtgtattga ccgattcctt gcggtccgaa tgggccgaac
     6781 ccgctcgtct ggctaagatc ggccgcagcg atcgcatcca tggcctccgc gaccggctgc
     6841 agaacagcgg gcagttcggt ttcaggcagg tcttgcaacg tgacaccctg tgcacggcgg
     6901 gagatgcaat aggtcaggct ctcgctgaat tccccaatgt caagcacttc cggaatcggg
     6961 agcgcggccg atgcaaagtg ccgataaaca taacgatctt tgtagaaacc atcggcgcag
     7021 ctatttaccc gcaggacata tccacgccct cctacatcga agctgaaagc acgagattct
     7081 tcgccctccg agagctgcat caggtcggag acgctgtcga acttttcgat cagaaacttc
     7141 tcgacagacg tcgcggtgag ttcaggcttt ttcatggtgg cggctggatc ggtcgaaagg
     7201 cccggagatg aggaagagga gaacagcgcg gcagacgtgc gcttttgaag cgtgcgagaa
     7261 tgccgggcct ccggaggacc ttcgggcgcc cgccccgccc ctgagcccgc ccctgagccc
     7321 gcccccggac ccaccccttc ccagcctctg agcccagaaa gcgaaggagc aaagctgcta
     7381 ttggccgctg ccccaaaggc ctacccgctt ccattgctca gcggtgctgt ccatctgcac
     7441 gagactagtg agacgtgcta cttccatttg tcacgtcctg cacgacgcga gctgcggggc
     7501 gggggggaac ttcctgacta ggggaggagt agaaggtggc gcgaaggggc caccaaagaa
     7561 cggagccggt tggcgcctac cggtggatgt ggaatgtgtg cgaggccaga ggccacttgt
     7621 gtagcgccaa gtgccagcgg ggctgctaaa gcgcatgctc cagactgcct tgggaaaagc
     7681 gcctccccta cccggtagaa ttcatcgatg atatcataac ttcgtataat gtatgctata
     7741 cgaagttatg cggccctcga ggtc
//
