LOCUS       sequence-14265-e        6947 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 6947)
  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 6947)
  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-68_HNF4A2
FEATURES             Location/Qualifiers
     source          1..6947
                     /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        235..286
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     primer_bind     237..257
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    288..306
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    309..327
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     337..361
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             527..556
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             566..595
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             605..634
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             644..673
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             683..712
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             746..775
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2899..2916)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2905..3129
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3262..3281)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3413..3460
                     /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             3468..4490
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3468..4490
                     /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    4620..4741
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4657..4676)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4711..4730
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4790..4806)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4790..4806)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4803..4825)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4814..4830
                     /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(4838..4844,4845..4862,4863..4868))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4883..4904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5021..5038)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5192..5780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5272..5291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5951..6742,6743..6811))
                     /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     6574..6593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6812..6916)
                     /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 cgttgacgca
      241 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga
      301 tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat
      361 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
      421 cctccggact ctagcgttta aacttaagct tggtaccgag ctcggatcca ctagtccagt
      481 gtggtggaat tctgcagata tccagcacag tggcggccgc gctatggagc aaaagctcat
      541 ttctgaagag gacttgaatg aaatggagca aaagctcatt tctgaagagg acttgaatga
      601 aatggagcaa aagctcattt ctgaagagga cttgaatgaa atggagcaaa agctcatttc
      661 tgaagaggac ttgaatgaaa tggagcaaaa gctcatttct gaagaggact tgaatgaaat
      721 ggagagcttg ggcgacctca ccatggagca aaagctcatt tctgaagagg acttgaattc
      781 aaggcctggt accatgcgac tctccaaaac cctcgtcgac atggacatgg ccgactacag
      841 tgctgcactg gacccagcct acaccaccct ggaatttgag aatgtgcagg tgttgacgat
      901 gggcaatgac acgtccccat cagaaggcac caacctcaac gcgcccaaca gcctgggtgt
      961 cagcgccctg tgtgccatct gcggggaccg ggccacgggc aaacactacg gtgcctcgag
     1021 ctgtgacggc tgcaagggct tcttccggag gagcgtgcgg aagaaccaca tgtactcctg
     1081 cagatttagc cggcagtgcg tggtggacaa agacaagagg aaccagtgcc gctactgcag
     1141 gctcaagaaa tgcttccggg ctggcatgaa gaaggaagcc gtccagaatg agcgggaccg
     1201 gatcagcact cgaaggtcaa gctatgagga cagcagcctg ccctccatca atgcgctcct
     1261 gcaggcggag gtcctgtccc gacagatcac ctcccccgtc tccgggatca acggcgacat
     1321 tcgggcgaag aagattgcca gcatcgcaga tgtgtgtgag tccatgaagg agcagctgct
     1381 ggttctcgtt gagtgggcca agtacatccc agctttctgc gagctccccc tggacgacca
     1441 ggtggccctg ctcagagccc atgctggcga gcacctgctg ctcggagcca ccaagagatc
     1501 catggtgttc aaggacgtgc tgctcctagg caatgactac attgtccctc ggcactgccc
     1561 ggagctggcg gagatgagcc gggtgtccat acgcatcctt gacgagctgg tgctgccctt
     1621 ccaggagctg cagatcgatg acaatgagta tgcctacctc aaagccatca tcttctttga
     1681 cccagatgcc aaggggctga gcgatccagg gaagatcaag cggctgcgtt cccaggtgca
     1741 ggtgagcttg gaggactaca tcaacgaccg ccagtatgac tcgcgtggcc gctttggaga
     1801 gctgctgctg ctgctgccca ccttgcagag catcacgtgg cagatgatcg agcagatcca
     1861 gttcatcaag ctcttcggca tggccaagat tgacaacctg ttgcaggaga tgctgctggg
     1921 agggtccccc agcgatgcac cccatgccca ccaccccctg caccctcacc tgatgcagga
     1981 acatatggga accaacgtca tcgttgccaa cacaatgccc actcacctca gcaacggaca
     2041 gatgtgtgag tggccccgac ccaggggaca ggcagccacc cctgagaccc cacagccctc
     2101 accgccaggt ggctcagggt ctgagcccta taagctcctg ccgggagccg tcgccacaat
     2161 cgtcaagccc ctctctgcca tcccccagcc gaccatcacc aagcaggaag ttatctagca
     2221 agccgctggg gcttgggggc tccactggct ccccccagcc ccctaagaga gcacctggtg
     2281 atcacgtggt cacggcaaag gaagacgtga tgccaggacc agtcccagag caggaatggg
     2341 aaggatgaag ggcccgagaa catggcctaa gggccacatc ccactgccac ccttgacgcc
     2401 ctgctctgga taacaagact ttgacttggg gagacctcta ctgccttgga caacttttct
     2461 catgttgaag ccactgcctt caccttcacc ttcatccatg tccaaccccc gacttcatcc
     2521 caaaggacag ccgcctggag atgacttgag gccttactta aacccagctc ccttcttccc
     2581 tagcctggtg cttctcctct cctagccccg gtcatggtgt ccagacagag ccctgtgagg
     2641 ctgggtccaa ttgtggcact tggggcacct tgctcctcct tctgctgctg cccccacctc
     2701 tgctgcctcc ctctgctgtc accttgctca gccatcccgt cttctccaac accacctcta
     2761 cagaggccaa ggaggccttg gaaacgattc ccccagtcat tctgggaaca tgttgtaagc
     2821 actgactggg accaggcacc aggcagggtc tagagcggcc gctcgagtct agagggcccg
     2881 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2941 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3001 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3061 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3121 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3181 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3241 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3301 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3361 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc tagaagttcc
     3421 tattccgaag ttcctattct ctagaaagta taggaacttc cttggccaaa aagcctgaac
     3481 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
     3541 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     3601 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     3661 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga
     3721 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
     3781 ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg
     3841 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     3901 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     3961 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     4021 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     4081 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
     4141 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     4201 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata
     4261 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
     4321 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
     4381 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
     4441 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtactac
     4501 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4561 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4621 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4681 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4741 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4801 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4861 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4921 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4981 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5041 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5101 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5161 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5221 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5281 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5341 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5401 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5461 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5521 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5581 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5641 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5701 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5761 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5821 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5881 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5941 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6001 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6061 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6121 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6181 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6241 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6301 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6361 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6421 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6481 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6541 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6601 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6661 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6721 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6781 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6841 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6901 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
