LOCUS       sequence-14264-e        7015 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 7015)
  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 7015)
  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_HNF4A2
FEATURES             Location/Qualifiers
     source          1..7015
                     /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(2967..2984)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2973..3197
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3330..3349)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3481..3528
                     /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             3536..4558
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3536..4558
                     /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    4688..4809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4725..4744)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4779..4798
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4858..4874)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4858..4874)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4871..4893)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4882..4898
                     /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(4906..4912,4913..4930,4931..4936))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4951..4972
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5089..5106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5260..5848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5340..5359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6019..6810,6811..6879))
                     /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     6642..6661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6880..6984)
                     /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 catgcgactc tccaaaaccc tcgtcgacat ggacatggcc
      901 gactacagtg ctgcactgga cccagcctac accaccctgg aatttgagaa tgtgcaggtg
      961 ttgacgatgg gcaatgacac gtccccatca gaaggcacca acctcaacgc gcccaacagc
     1021 ctgggtgtca gcgccctgtg tgccatctgc ggggaccggg ccacgggcaa acactacggt
     1081 gcctcgagct gtgacggctg caagggcttc ttccggagga gcgtgcggaa gaaccacatg
     1141 tactcctgca gatttagccg gcagtgcgtg gtggacaaag acaagaggaa ccagtgccgc
     1201 tactgcaggc tcaagaaatg cttccgggct ggcatgaaga aggaagccgt ccagaatgag
     1261 cgggaccgga tcagcactcg aaggtcaagc tatgaggaca gcagcctgcc ctccatcaat
     1321 gcgctcctgc aggcggaggt cctgtcccga cagatcacct cccccgtctc cgggatcaac
     1381 ggcgacattc gggcgaagaa gattgccagc atcgcagatg tgtgtgagtc catgaaggag
     1441 cagctgctgg ttctcgttga gtgggccaag tacatcccag ctttctgcga gctccccctg
     1501 gacgaccagg tggccctgct cagagcccat gctggcgagc acctgctgct cggagccacc
     1561 aagagatcca tggtgttcaa ggacgtgctg ctcctaggca atgactacat tgtccctcgg
     1621 cactgcccgg agctggcgga gatgagccgg gtgtccatac gcatccttga cgagctggtg
     1681 ctgcccttcc aggagctgca gatcgatgac aatgagtatg cctacctcaa agccatcatc
     1741 ttctttgacc cagatgccaa ggggctgagc gatccaggga agatcaagcg gctgcgttcc
     1801 caggtgcagg tgagcttgga ggactacatc aacgaccgcc agtatgactc gcgtggccgc
     1861 tttggagagc tgctgctgct gctgcccacc ttgcagagca tcacgtggca gatgatcgag
     1921 cagatccagt tcatcaagct cttcggcatg gccaagattg acaacctgtt gcaggagatg
     1981 ctgctgggag ggtcccccag cgatgcaccc catgcccacc accccctgca ccctcacctg
     2041 atgcaggaac atatgggaac caacgtcatc gttgccaaca caatgcccac tcacctcagc
     2101 aacggacaga tgtgtgagtg gccccgaccc aggggacagg cagccacccc tgagacccca
     2161 cagccctcac cgccaggtgg ctcagggtct gagccctata agctcctgcc gggagccgtc
     2221 gccacaatcg tcaagcccct ctctgccatc ccccagccga ccatcaccaa gcaggaagtt
     2281 atctagcaag ccgctggggc ttgggggctc cactggctcc ccccagcccc ctaagagagc
     2341 acctggtgat cacgtggtca cggcaaagga agacgtgatg ccaggaccag tcccagagca
     2401 ggaatgggaa ggatgaaggg cccgagaaca tggcctaagg gccacatccc actgccaccc
     2461 ttgacgccct gctctggata acaagacttt gacttgggga gacctctact gccttggaca
     2521 acttttctca tgttgaagcc actgccttca ccttcacctt catccatgtc caacccccga
     2581 cttcatccca aaggacagcc gcctggagat gacttgaggc cttacttaaa cccagctccc
     2641 ttcttcccta gcctggtgct tctcctctcc tagccccggt catggtgtcc agacagagcc
     2701 ctgtgaggct gggtccaatt gtggcacttg gggcaccttg ctcctccttc tgctgctgcc
     2761 cccacctctg ctgcctccct ctgctgtcac cttgctcagc catcccgtct tctccaacac
     2821 cacctctaca gaggccaagg aggccttgga aacgattccc ccagtcattc tgggaacatg
     2881 ttgtaagcac tgactgggac caggcaccag gcagggtcta gagcggccgc tcgagtctag
     2941 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3001 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3061 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3121 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3181 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3241 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3301 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3361 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3421 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaccta
     3481 gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcct tggccaaaaa
     3541 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     3601 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     3661 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     3721 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt
     3781 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     3841 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
     3901 tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     3961 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     4021 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     4081 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     4141 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     4201 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     4261 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     4321 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     4381 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     4441 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     4501 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca
     4561 cgtactacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4621 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4681 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4741 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4801 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4861 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4921 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4981 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5041 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5101 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5161 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5221 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5281 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5341 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5401 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5461 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5521 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5581 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5641 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5701 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5761 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5821 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5881 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5941 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6001 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6061 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6121 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6181 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6241 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6301 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6361 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6421 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6481 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6541 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6601 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6661 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6721 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6781 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6841 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6901 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6961 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
