LOCUS       sequence-14263-e        7096 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 7096)
  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 7096)
  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-218_HNF4A2
FEATURES             Location/Qualifiers
     source          1..7096
                     /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        232..435
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     386..406
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    437..455
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    458..476
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     486..510
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             676..705
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             715..744
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             754..783
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             793..822
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             832..861
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             895..924
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3048..3065)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3054..3278
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3411..3430)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3562..3609
                     /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             3617..4639
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3617..4639
                     /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    4769..4890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4806..4825)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4860..4879
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4939..4955)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4939..4955)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4952..4974)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4963..4979
                     /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(4987..4993,4994..5011,5012..5017))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5032..5053
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5170..5187)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5341..5929)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5421..5440)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6100..6891,6892..6960))
                     /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     6723..6742
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6961..7065)
                     /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 cgtgatgcgg
      241 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      301 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      361 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      421 tatataagca gagctctccc tatcagtgat agagatctcc ctatcagtga tagagatcgt
      481 cgacgagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac
      541 ctccatagaa gacaccggga ccgatccagc ctccggactc tagcgtttaa acttaagctt
      601 ggtaccgagc tcggatccac tagtccagtg tggtggaatt ctgcagatat ccagcacagt
      661 ggcggccgcg ctatggagca aaagctcatt tctgaagagg acttgaatga aatggagcaa
      721 aagctcattt ctgaagagga cttgaatgaa atggagcaaa agctcatttc tgaagaggac
      781 ttgaatgaaa tggagcaaaa gctcatttct gaagaggact tgaatgaaat ggagcaaaag
      841 ctcatttctg aagaggactt gaatgaaatg gagagcttgg gcgacctcac catggagcaa
      901 aagctcattt ctgaagagga cttgaattca aggcctggta ccatgcgact ctccaaaacc
      961 ctcgtcgaca tggacatggc cgactacagt gctgcactgg acccagccta caccaccctg
     1021 gaatttgaga atgtgcaggt gttgacgatg ggcaatgaca cgtccccatc agaaggcacc
     1081 aacctcaacg cgcccaacag cctgggtgtc agcgccctgt gtgccatctg cggggaccgg
     1141 gccacgggca aacactacgg tgcctcgagc tgtgacggct gcaagggctt cttccggagg
     1201 agcgtgcgga agaaccacat gtactcctgc agatttagcc ggcagtgcgt ggtggacaaa
     1261 gacaagagga accagtgccg ctactgcagg ctcaagaaat gcttccgggc tggcatgaag
     1321 aaggaagccg tccagaatga gcgggaccgg atcagcactc gaaggtcaag ctatgaggac
     1381 agcagcctgc cctccatcaa tgcgctcctg caggcggagg tcctgtcccg acagatcacc
     1441 tcccccgtct ccgggatcaa cggcgacatt cgggcgaaga agattgccag catcgcagat
     1501 gtgtgtgagt ccatgaagga gcagctgctg gttctcgttg agtgggccaa gtacatccca
     1561 gctttctgcg agctccccct ggacgaccag gtggccctgc tcagagccca tgctggcgag
     1621 cacctgctgc tcggagccac caagagatcc atggtgttca aggacgtgct gctcctaggc
     1681 aatgactaca ttgtccctcg gcactgcccg gagctggcgg agatgagccg ggtgtccata
     1741 cgcatccttg acgagctggt gctgcccttc caggagctgc agatcgatga caatgagtat
     1801 gcctacctca aagccatcat cttctttgac ccagatgcca aggggctgag cgatccaggg
     1861 aagatcaagc ggctgcgttc ccaggtgcag gtgagcttgg aggactacat caacgaccgc
     1921 cagtatgact cgcgtggccg ctttggagag ctgctgctgc tgctgcccac cttgcagagc
     1981 atcacgtggc agatgatcga gcagatccag ttcatcaagc tcttcggcat ggccaagatt
     2041 gacaacctgt tgcaggagat gctgctggga gggtccccca gcgatgcacc ccatgcccac
     2101 caccccctgc accctcacct gatgcaggaa catatgggaa ccaacgtcat cgttgccaac
     2161 acaatgccca ctcacctcag caacggacag atgtgtgagt ggccccgacc caggggacag
     2221 gcagccaccc ctgagacccc acagccctca ccgccaggtg gctcagggtc tgagccctat
     2281 aagctcctgc cgggagccgt cgccacaatc gtcaagcccc tctctgccat cccccagccg
     2341 accatcacca agcaggaagt tatctagcaa gccgctgggg cttgggggct ccactggctc
     2401 cccccagccc cctaagagag cacctggtga tcacgtggtc acggcaaagg aagacgtgat
     2461 gccaggacca gtcccagagc aggaatggga aggatgaagg gcccgagaac atggcctaag
     2521 ggccacatcc cactgccacc cttgacgccc tgctctggat aacaagactt tgacttgggg
     2581 agacctctac tgccttggac aacttttctc atgttgaagc cactgccttc accttcacct
     2641 tcatccatgt ccaacccccg acttcatccc aaaggacagc cgcctggaga tgacttgagg
     2701 ccttacttaa acccagctcc cttcttccct agcctggtgc ttctcctctc ctagccccgg
     2761 tcatggtgtc cagacagagc cctgtgaggc tgggtccaat tgtggcactt ggggcacctt
     2821 gctcctcctt ctgctgctgc ccccacctct gctgcctccc tctgctgtca ccttgctcag
     2881 ccatcccgtc ttctccaaca ccacctctac agaggccaag gaggccttgg aaacgattcc
     2941 cccagtcatt ctgggaacat gttgtaagca ctgactggga ccaggcacca ggcagggtct
     3001 agagcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     3061 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3121 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3181 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3241 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3301 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3361 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3421 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3481 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3541 gggtgatggt tcacgtacct agaagttcct attccgaagt tcctattctc tagaaagtat
     3601 aggaacttcc ttggccaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     3661 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     3721 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     3781 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     3841 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     3901 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     3961 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     4021 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     4081 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     4141 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     4201 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     4261 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     4321 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     4381 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     4441 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     4501 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     4561 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     4621 tccgagggca aaggaatagc acgtactacg agatttcgat tccaccgccg ccttctatga
     4681 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4741 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4801 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4861 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4921 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4981 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5041 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5101 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5161 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5221 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5281 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5341 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5401 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5461 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5521 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5581 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5641 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5701 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5761 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5821 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5881 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5941 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6001 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6061 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6121 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6181 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6241 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6301 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6361 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6421 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6481 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6541 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6601 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6661 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6721 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6781 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6841 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6901 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6961 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7021 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7081 agtgccacct gacgtc
//
