LOCUS       sequence-137732-        5265 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 5265)
  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 5265)
  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: pCMV-HA GCNF
FEATURES             Location/Qualifiers
     source          1..5265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     531..555
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          672..768
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             832..858
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2404..2538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2428..2447)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2482..2501
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2649..2665)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2649..2665)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2662..2684)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2673..2689
                     /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(2697..2703,2704..2721,2722..2727))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2742..2763
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2880..2897)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3051..3639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3131..3150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3810..4601,4602..4670))
                     /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     4433..4452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4671..4775)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4843..4861
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4899..4921)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5021..5040
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5234..5256
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5248..5265
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5249..5265
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag
      661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat gtacccatac
      841 gatgttccag attacgctct tatggccatg gaggcccgaa ttcgcatgga gcgggacgaa
      901 cggccaccta gcggaggggg aggcggcggg ggctcggcgg ggttcctgga gccgcccgcc
      961 gcgctccctc cgccgccgcg cagcggtttc tgtcaggatg aattggcaga gcttgatcca
     1021 ggcactaatg gagagactga cagtttaaca cttggccaag gccatatacc tgtttccgtc
     1081 ccagatgatc gagctgaaca acgaacctgt ctcatctgtg gggaccgcgc tacgggcttg
     1141 cactatggga tcatctcctg cgagggctgc aaggggtttt tcaagaggag catttgcaac
     1201 aaacgggtgt atcggtgcag tcgtgacaag aactgtgtca tgtcccggaa gcagaggaac
     1261 agatgtcagt actgccgcct gctcaagtgt ctccagatgg gcatgaacag gaaggctatc
     1321 agagaagatg gcatgcctgg aggccggaac aagagcattg gaccagtcca gatatcagaa
     1381 gaagaaattg aaagaatcat gtctggacag gagtttgagg aagaagccaa tcactggagc
     1441 aaccatggtg acagcgacca cagttcccct gggaacaggg cttcagagag caaccagccc
     1501 tcaccaggct ccacactatc atccagtagg tctgtggaac taaatggatt catggcattc
     1561 agggatcagt acatggggat gtcagtgcct ccacattatc aatacatacc acaccttttt
     1621 agctattctg gccactcacc acttttgccc ccacaagctc gaagcctgga ccctcagtcc
     1681 tacagtctga ttcatcagct gatgtcagcc gaagacctgg agccattggg cacacctatg
     1741 ttgattgaag atgggtatgc tgtgacacag gcagaactgt ttgctctgct ttgccgcctg
     1801 gccgacgagt tgctctttag gcagattgcc tggatcaaga agctgccttt cttctgcgag
     1861 ctctcaatca aggattacac gtgcctcttg agctctacgt ggcaggagtt aatcctgctc
     1921 tcctccctca cagtgtacag caagcagatc tttggggagc tggctgatgt cacagccaag
     1981 tactcaccct ctgatgaaga actccacaga tttagtgatg aagggatgga ggtgattgaa
     2041 cgactcatct acctatatca caagttccat cagctgaagg tcagcaacga ggagtacgca
     2101 tgcatgaaag caattaactt cctggatcaa gatatcaggg gtctgaccag tgcctcacag
     2161 ctggaacaac tgaacaagcg gtattggtac atttgtcagg atttcactga atataaatac
     2221 acacatcagc caaaccgctt tcctgatctt atgatgtgct tgccagagat ccgatacatc
     2281 gcaggcaaga tggtgaatgt gcccctggag cagctgcccc tcctctttaa ggtggtgctg
     2341 cactcctgca agacaagtac ggtgaaggag tgacctgtgc cctcgaggta ccgcggccgc
     2401 ggggatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     2461 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2521 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2581 gaggtgtggg aggttttttc ggatcctcta gagtcgatct gcaggcatgc tagcttggcg
     2641 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2701 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2761 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2821 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2881 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2941 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3001 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3061 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3121 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3181 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3241 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3301 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3361 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3421 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3481 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3541 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3601 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3661 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3721 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3781 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3841 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3901 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3961 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4021 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4081 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4141 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4201 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4261 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4321 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4381 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4441 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4501 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4561 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4621 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4681 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4741 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4801 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     4861 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     4921 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     4981 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     5041 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     5101 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     5161 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     5221 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagt
//
