LOCUS       sequence-147143-        7800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of zNanog Homeodomain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7800)
  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 7800)
  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-zNanogHD-pgk-hph
FEATURES             Location/Qualifiers
     source          1..7800
                     /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(2031..2055)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2076..2092)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2100..2116
                     /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(2124..2130,2131..2148,2149..2154))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2169..2190
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2250..2267)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2256..2480
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    2557..2590
                     /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(2813..2832)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2814..2832)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2842..2859)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2842..2858)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2851..2873)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3000..3455
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3087..3106)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3297..3318
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3481..3585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3586..3654,3655..4446)
                     /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(3804..3823)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4617..5205
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5106..5125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5359..5376
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5493..5514
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5529..5534,5535..5552,5553..5559)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5567..5583
                     /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     5591..5607
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5626..5646
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5628..5646
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   5669..5703
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     CDS             complement(6186..7211)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(6186..7211)
                     /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(7230..7729)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     7671..7694
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    7752..7785
                     /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 cggcctccga ccctgccccg
     1801 caagacccgg gctgcttttt ctgaagaaca gatgaatgct ttggtcaatc ggtttaatgt
     1861 ccagagatac ctcacgcctg cagagatgaa aacgcttgct ggagcaacag gattaacata
     1921 caagcaggta aaaacatggt ttcagaaccg caggatgaaa ctcaagaggc atcagagaga
     1981 cagcagttgg atgactgaga gatatgttgt caatgctgta cccaactaag acccagcttt
     2041 cttgtacaaa gtggtgatag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     2101 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     2161 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2221 tcgggaaacc tgtcgtgcca gcggatcagc ctcgactgtg ccttctagtt gccagccatc
     2281 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2341 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2401 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2461 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct cgagatccac
     2521 tagttctagc ctcgaggcta gagcggccgc gtcgagataa cttcgtataa tgtatgctat
     2581 acgaagttat gttttgttac tttatagaag aaattttgag tttttgtttt tttttaataa
     2641 ataaataaac ataaataaat tgtttgttga atttattatt agtatgtaag tgtaaatata
     2701 ataaaactta atatctattc aaattaataa ataaacgtca attttacgca tgattatctt
     2761 taacgtacgt cacaatatga ttatctttct agggttaaag agctccaatt cgccctatag
     2821 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     2881 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     2941 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     3001 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3061 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3121 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3181 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3241 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3301 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3361 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3421 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     3481 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3541 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3601 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3661 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3721 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3781 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3841 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3901 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3961 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4021 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4081 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4141 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4201 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4261 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4321 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4381 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4441 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4501 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4561 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4621 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4681 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4741 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     4801 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4861 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4921 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4981 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5041 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5101 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5161 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5221 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5281 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5341 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     5401 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     5461 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     5521 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     5581 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     5641 taaagggaac aaaagctggg taccttaacc ctagaaagat agtctgcgta aaattgacgc
     5701 atgcattctt gaaatattgc tctctctttc taaatagcgc gaatccgtcg ctgtgcattt
     5761 aggacatctc agtcgccgct tggagctccc gtgaggcgtg cttgtcaatg cggtaagtgt
     5821 cactgatttt gaactataac gaccgcgtga gtcaaaatga cgcatgatta tcttttacgt
     5881 gacttttaag atttaactca tacgataatt atattgttat ttcatgttct acttacgtga
     5941 taacttatta tatatatatt ttcttgttat agatatccaa ctatgaaaca ttatcataaa
     6001 gcaatagtaa agagccttca gtaaagagca ggcatttatc taatcccacc ccacccccac
     6061 ccccgtagct ccaatccttc cattcaaaat gtaggtactc tgttctcacc cttcttaaca
     6121 aagtatgaca ggaaaaaact tccatgtgga catctttatt gtttaataga tcatcaattt
     6181 ctcgactatt cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt
     6241 acttctacac agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg
     6301 acagtcccgg ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca
     6361 tcgaaattgc cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac
     6421 gcccggagcc gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc
     6481 tgctccatac aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa
     6541 tccccgaaca tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg
     6601 acattgttgg agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc
     6661 caaagcatca gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca
     6721 gtttgccagt gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg
     6781 tattgaccga ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcc
     6841 gcagcgatcg catccatggc ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca
     6901 ggcaggtctt gcaacgtgac accctgtgca cggcgggaga tgcaataggt caggctctcg
     6961 ctgaattccc caatgtcaag cacttccgga atcgggagcg cggccgatgc aaagtgccga
     7021 taaacataac gatctttgta gaaaccatcg gcgcagctat ttacccgcag gacatatcca
     7081 cgccctccta catcgaagct gaaagcacga gattcttcgc cctccgagag ctgcatcagg
     7141 tcggagacgc tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc ggtgagttca
     7201 ggctttttca tggtggcggc tggatcggtc gaaaggcccg gagatgagga agaggagaac
     7261 agcgcggcag acgtgcgctt ttgaagcgtg cgagaatgcc gggcctccgg aggaccttcg
     7321 ggcgcccgcc ccgcccctga gcccgcccct gagcccgccc ccggacccac cccttcccag
     7381 cctctgagcc cagaaagcga aggagcaaag ctgctattgg ccgctgcccc aaaggcctac
     7441 ccgcttccat tgctcagcgg tgctgtccat ctgcacgaga ctagtgagac gtgctacttc
     7501 catttgtcac gtcctgcacg acgcgagctg cggggcgggg gggaacttcc tgactagggg
     7561 aggagtagaa ggtggcgcga aggggccacc aaagaacgga gccggttggc gcctaccggt
     7621 ggatgtggaa tgtgtgcgag gccagaggcc acttgtgtag cgccaagtgc cagcggggct
     7681 gctaaagcgc atgctccaga ctgccttggg aaaagcgcct cccctacccg gtagaattca
     7741 tcgatgatat cataacttcg tataatgtat gctatacgaa gttatgcggc cctcgaggtc
//
