LOCUS       sequence-163190-        7397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7397)
  AUTHORS   Gu P, LeMenuet D, Chung AC, Mancini M, Wheeler DA, Cooney AJ
  TITLE     Orphan nuclear receptor GCNF is required for the repression of 
            pluripotency genes during retinoic acid-induced embryonic stem cell 
            differentiation.
  JOURNAL   Mol Cell Biol. 2005 Oct . 25(19):8507-19.
  PUBMED    16166633
REFERENCE   2  (bases 1 to 7397)
  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: "Mol Cell 
            Biol. 2005 Oct . 25(19):8507-19."
COMMENT     Sequence Label: Nanog5P reporter
FEATURES             Location/Qualifiers
     source          1..7397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1005..1014
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2609..2618
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2615..4267
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(2681..2699)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4308..4429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4319..4338)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4373..4392
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4729..4746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4900..5488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4980..4999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5659..6450,6451..6519))
                     /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     6282..6301
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6520..6624)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6651..7106
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6738..6757)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6948..6969
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7237..7285
                     /note="synthetic polyadenylation signal"
     misc_feature    7299..7390
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7339..7358
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccacta agggcgaatt ccagcacact ggcggccgtt actagtggat ccgagctcgg
       61 taccaacagt gggtctgaag ccttcgaggg aggagggcaa gatgccgcgc tctgcgtttc
      121 tccagctgga tgtggacaga ggtcaaccag ccacattagt ttatgtctaa aagtgtctaa
      181 ttgaaacaag aaatggctgc tttagccggg tgtggtggca cacgccttta atcccagcac
      241 tcaggaggca gaggcaggca gatttctgag ttcaaggcca gcctggtcta cagagtgagt
      301 tccaggacag ccagagctac acagagaaac cctgtctcga aaaaccaacc aaccaaccaa
      361 ccaaacaaac aaacaaaaaa ggaaatggct ggtttaatta tatcacactg ttcatcagaa
      421 taaagagaaa aatctcaaaa aaaaaaaaaa agaggttcag tcaggctggg caatggaggc
      481 agtagtggcc gtggtggtct tggcagcagt ggaggccttg gtggcagtgg tgacagacag
      541 tgatagtgtg gcagcagttg aggcagtggt ggtcgtagtg gcagtgatgg cattggtggc
      601 agggttggtg tggctcagag gtggcagtgg tggcagtgga ggcattgcag gctttagtgg
      661 caatggtagt ggggcagcag tggtagccat ggtggcacag tggtagcaac ggtggtagtg
      721 gcagccgtgg tggccgcaga ggcagtggta acaatggtgg cttggtggca gtgctgacag
      781 tggtagtatt ggtagcattg gtggtagtgg aagaagggaa gtggtggtaa tggtggcagg
      841 ggtggcagca gaggcagaca gtggtgacga tggtggcagt ggtgctgtgg cacagtggtg
      901 gcattggtgg catggcacag tggtggcaat gatggtgtgg tgcagtggta gcagtggaga
      961 cagtggtggc agtggagaca gtagtagtat ggtggctgtg gtggccatgg cggcagtggt
     1021 ggcacataca ggcctttaat gccagcacat gggaggcaga ggcagttggc tctctgtgag
     1081 ttggaagcca gcttgttcta cagagcaagt cccaggacag ccagggctac acagagaaac
     1141 cctgcctaga ataataacaa aactcaaaca aacaccacca acccccgcgc tccgccccgc
     1201 cccccccaaa aaaaacccca aaccaaaaag agccattcaa gcttagtctt ttgtagaata
     1261 aaacccagga agaaccactc ctaccaatac tcactgtggt agagtcttca catgggagat
     1321 gttacgagag gacggccctt ccctctctgc ttatacacag aagccgactt aagctgggtt
     1381 agagtgcttt cactcactta tctgtgagca caaggactga tcggcaaact ttgaacttgg
     1441 gatgtggaaa taggaagatc aggagtttgg gaccagctag agcaattgac tgtgaggcca
     1501 gtctcgggta cgtgaaactc tatctcaaag aaatgaccaa aaccaaattt gaaatacgat
     1561 aaatttcttc ttccattgct tagacggctg aggcacttgg gattgcagta agtctgaagc
     1621 cagcctaggt tttacagtga gaacttgtct caaaacaaaa cagaaggcct ctacattaat
     1681 ttaaacactc cttaaattgg gcatggtggt agacaagcct ggtctacata ctgagtataa
     1741 gctactcaag gcaacagaga aaaacctgtc tcaaaaccaa agcatggacc aacttactaa
     1801 ggtagcccga gtcttaagca ggacacgggc tctttcttca gacttgcgtt aaaaagccgc
     1861 acttttggag ggaagatttc cccaggtttc ccaatgtgaa gagcaagcaa gaaacgctga
     1921 gtgctgaaag gaaagccgtg tataaacaga gactttggca gcaaggtctg actctttcat
     1981 gtctgtagaa agaatggaag aggaaactca gatcccccac ttgacctgaa acttcccact
     2041 agagatcgcc agggtctgga ggtgcagccg tggttaaaag atgaataaag tgaaatgagg
     2101 taaagcctct tttcgggggg gggggatgtc tttagatcag aggatgcccc ctaagctttc
     2161 cctccctccc agtctgggtc accttacagc ttcttttgca ttacaatgtc catggtggac
     2221 cctgcaggtg ggattaactg tgaattcaca gggctggtgg ggcgtgggtg ccgcctgggt
     2281 gcctgggaga atagggggtg ggtagggtag gaggcttgag gggggaggag caggacctac
     2341 cctttaaatc tatcgccttg agccgttggc cttcagatag gctgatttgg ttggtgtctt
     2401 gctctttctg tgggaaggct gcggctcact tccttctgac ttcttgataa ttttgcatta
     2461 gacatttaac tcttctttct gtgatctttc cttctagaca ctgagttttt tggttgttgc
     2521 ctaaaacctt ttcagaaatc ccttccctcg ccatcacact cgagatctgc gatctaagta
     2581 agcttggcat tccggtactg ttggtaaagc caccatggaa gacgccaaaa acataaagaa
     2641 aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac tgcataaggc
     2701 tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt
     2761 ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg
     2821 atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt
     2881 tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta
     2941 taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg tgttcgtttc
     3001 caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat
     3061 tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac
     3121 atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct tcgataggga
     3181 caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta aaggtgtcgc
     3241 tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta tttttggcaa
     3301 tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat
     3361 gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga
     3421 agaagagctg tttctgagga gccttcagga ttacaagatt caaagtgcgc tgctggtgcc
     3481 aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt
     3541 acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag cggttgccaa
     3601 gaggttccat ctgccaggta tcaggcaagg atatgggctc actgagacta catcagctat
     3661 tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt
     3721 tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc aaagaggcga
     3781 actgtgtgtg agaggtccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa
     3841 cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga
     3901 cgaacacttc ttcatcgttg accgcctgaa gtctctgatt aagtacaaag gctatcaggt
     3961 ggctcccgct gaattggaat ccatcttgct ccaacacccc aacatcttcg acgcaggtgt
     4021 cgcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca
     4081 cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc
     4141 gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa
     4201 actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagatcgc
     4261 cgtgtaattc tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga
     4321 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     4381 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     4441 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     4501 aaacctctac aaatgtggta aaatcgataa ggatccgtcg accgatgccc ttgagagcct
     4561 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     4621 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctttat gactgtcttc
     4681 tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga
     4741 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4801 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4861 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4921 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4981 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5041 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5101 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5161 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5221 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5281 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5341 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5401 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5461 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5521 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5581 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5641 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5701 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5761 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5821 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5881 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5941 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6001 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6061 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6121 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6181 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6241 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6301 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6361 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6421 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6481 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6541 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6601 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg
     6661 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     6721 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     6781 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     6841 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     6901 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     6961 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     7021 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     7081 taacaaaata ttaacgctta caatttgcca ttcgccattc aggctgcgca actgttggga
     7141 agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat gataagtaag
     7201 taatattaag gtacgggagg tacttggagc ggccgcaata aaatatcttt attttcatta
     7261 catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac
     7321 aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga
     7381 acatttctct atcgata
//
