LOCUS       sequence-14266-e        6949 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 6949)
  AUTHORS   Senkel S, Waldner C, Ryffel GU, Thomas H
  TITLE     Improved conditional expression systems resulting in physiological 
            level of HNF4alpha expression confirm HNF4alpha induced apoptosis in
            the pancreatic beta-cell line INS-1.
  JOURNAL   BMC Res Notes. 2009 Oct 17;2:210. doi: 10.1186/1756-0500-2-210.
  PUBMED    19835622
REFERENCE   2  (bases 1 to 6949)
  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: "BMC Res 
            Notes."; date: "2009-10-17"; volume: "2:210. doi"; pages: " 
            10.1186/1756-0500-2-210"
COMMENT     Sequence Label: CMV-138_HNF4A8
FEATURES             Location/Qualifiers
     source          1..6949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        255..354
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     305..325
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    356..374
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    377..395
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     405..429
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             595..624
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             634..663
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             673..702
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             712..741
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             751..780
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             814..843
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2901..2918)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2907..3131
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3264..3283)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3415..3462
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3470..4492
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3470..4492
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4622..4743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4659..4678)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4713..4732
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4792..4808)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4792..4808)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4805..4827)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4816..4832
                     /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(4840..4846,4847..4864,4865..4870))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4885..4906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5023..5040)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5194..5782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5274..5293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5953..6744,6745..6813))
                     /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     6576..6595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6814..6918)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtcaatggg
      241 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      301 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct
      361 atcagtgata gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac
      421 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
      481 cgatccagcc tccggactct agcgtttaaa cttaagcttg gtaccgagct cggatccact
      541 agtccagtgt ggtggaattc tgcagatatc cagcacagtg gcggccgcgc tatggagcaa
      601 aagctcattt ctgaagagga cttgaatgaa atggagcaaa agctcatttc tgaagaggac
      661 ttgaatgaaa tggagcaaaa gctcatttct gaagaggact tgaatgaaat ggagcaaaag
      721 ctcatttctg aagaggactt gaatgaaatg gagcaaaagc tcatttctga agaggacttg
      781 aatgaaatgg agagcttggg cgacctcacc atggagcaaa agctcatttc tgaagaggac
      841 ttgaattcaa ggcctggtac catggtcagc gtgaacgcgc ccctcggggc tccagtggag
      901 agttcttacg acacgtcccc atcagaaggc accaacctca acgcgcccaa cagcctgggt
      961 gtcagcgccc tgtgtgccat ctgcggggac cgggccacgg gcaaacacta cggtgcctcg
     1021 agctgtgacg gctgcaaggg cttcttccgg aggagcgtgc ggaagaacca catgtactcc
     1081 tgcagattta gccggcagtg cgtggtggac aaagacaaga ggaaccagtg ccgctactgc
     1141 aggctcaaga aatgcttccg ggctggcatg aagaaggaag ccgtccagaa tgagcgggac
     1201 cggatcagca ctcgaaggtc aagctatgag gacagcagcc tgccctccat caatgcgctc
     1261 ctgcaggcgg aggtcctgtc ccgacagatc acctcccccg tctccgggat caacggcgac
     1321 attcgggcga agaagattgc cagcatcgca gatgtgtgtg agtccatgaa ggagcagctg
     1381 ctggttctcg ttgagtgggc caagtacatc ccagctttct gcgagctccc cctggacgac
     1441 caggtggccc tgctcagagc ccatgctggc gagcacctgc tgctcggagc caccaagaga
     1501 tccatggtgt tcaaggacgt gctgctccta ggcaatgact acattgtccc tcggcactgc
     1561 ccggagctgg cggagatgag ccgggtgtcc atacgcatcc ttgacgagct ggtgctgccc
     1621 ttccaggagc tgcagatcga tgacaatgag tatgcctacc tcaaagccat catcttcttt
     1681 gacccagatg ccaaggggct gagcgatcca gggaagatca agcggctgcg ttcccaggtg
     1741 caggtgagct tggaggacta catcaacgac cgccagtatg actcgcgtgg ccgctttgga
     1801 gagctgctgc tgctgctgcc caccttgcag agcatcacgt ggcagatgat cgagcagatc
     1861 cagttcatca agctcttcgg catggccaag attgacaacc tgttgcagga gatgctgctg
     1921 ggagggtccc ccagcgatgc accccatgcc caccaccccc tgcaccctca cctgatgcag
     1981 gaacatatgg gaaccaacgt catcgttgcc aacacaatgc ccactcacct cagcaacgga
     2041 cagatgtgtg agtggccccg acccagggga caggcagcca cccctgagac cccacagccc
     2101 tcaccgccag gtggctcagg gtctgagccc tataagctcc tgccgggagc cgtcgccaca
     2161 atcgtcaagc ccctctctgc catcccccag ccgaccatca ccaagcagga agttatctag
     2221 caagccgctg gggcttgggg gctccactgg ctccccccag ccccctaaga gagcacctgg
     2281 tgatcacgtg gtcacggcaa aggaagacgt gatgccagga ccagtcccag agcaggaatg
     2341 ggaaggatga agggcccgag aacatggcct aagggccaca tcccactgcc acccttgacg
     2401 ccctgctctg gataacaaga ctttgacttg gggagacctc tactgccttg gacaactttt
     2461 ctcatgttga agccactgcc ttcaccttca ccttcatcca tgtccaaccc ccgacttcat
     2521 cccaaaggac agccgcctgg agatgacttg aggccttact taaacccagc tcccttcttc
     2581 cctagcctgg tgcttctcct ctcctagccc cggtcatggt gtccagacag agccctgtga
     2641 ggctgggtcc aattgtggca cttggggcac cttgctcctc cttctgctgc tgcccccacc
     2701 tctgctgcct ccctctgctg tcaccttgct cagccatccc gtcttctcca acaccacctc
     2761 tacagaggcc aaggaggcct tggaaacgat tcccccagtc attctgggaa catgttgtaa
     2821 gcactgactg ggaccaggca ccaggcaggg tctagagcgg ccgctcgagt ctagagggcc
     2881 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2941 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3001 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3061 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3121 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3181 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3241 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3301 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3361 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta cctagaagtt
     3421 cctattccga agttcctatt ctctagaaag tataggaact tccttggcca aaaagcctga
     3481 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     3541 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     3601 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     3661 gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga
     3721 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     3781 aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc
     3841 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     3901 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     3961 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     4021 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     4081 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     4141 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     4201 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta
     4261 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     4321 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     4381 gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact
     4441 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agcacgtact
     4501 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4561 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4621 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4681 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4741 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4801 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4861 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4921 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4981 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5041 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5101 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5161 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5221 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5281 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5341 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5401 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5461 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5521 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5581 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5641 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5701 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5761 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5821 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5881 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5941 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6001 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6061 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6121 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6181 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6241 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6301 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6361 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6421 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6481 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6541 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6601 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6661 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6721 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6781 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6841 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6901 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
