LOCUS       sequence-14275-e        7213 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 7213)
  AUTHORS   Thomas H, Senkel S, Erdmann S, Arndt T, Turan G, Klein-Hitpass L, 
            Ryffel GU
  TITLE     Pattern of genes influenced by conditional expression of the 
            transcription factors HNF6, HNF4alpha and HNF1beta in a pancreatic 
            beta-cell line.
  JOURNAL   Nucleic Acids Res. 2004  . 32(19):e150.
  PUBMED    15520459
REFERENCE   2  (bases 1 to 7213)
  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: "Nucleic 
            Acids Res. 2004  . 32(19):e150."
COMMENT     Sequence Label: FR_HNF1B_A263insGG
FEATURES             Location/Qualifiers
     source          1..7213
                     /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"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1031..1060
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1070..1099
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1109..1138
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1148..1177
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1187..1216
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1250..1279
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3165..3182)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3171..3395
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3528..3547)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3679..3726
                     /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             3734..4756
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3734..4756
                     /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    4886..5007
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4923..4942)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4977..4996
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5056..5072)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5056..5072)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5069..5091)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5080..5096
                     /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(5104..5110,5111..5128,5129..5134))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5149..5170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5287..5304)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5458..6046)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5538..5557)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6217..7008,7009..7077))
                     /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     6840..6859
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7078..7182)
                     /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 cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagttct aggatcgatt
     1021 taaagctatg gagcaaaagc tcatttctga agaggacttg aatgaaatgg agcaaaagct
     1081 catttctgaa gaggacttga atgaaatgga gcaaaagctc atttctgaag aggacttgaa
     1141 tgaaatggag caaaagctca tttctgaaga ggacttgaat gaaatggagc aaaagctcat
     1201 ttctgaagag gacttgaatg aaatggagag cttgggcgac ctcaccatgg agcaaaagct
     1261 catttctgaa gaggacttga attcaaggcc tctcgaagct tccatggtgt ccaagctcac
     1321 gtcgctccag caagaactcc tgagcgccct gctgagctcc ggggtcacca aggaggtgct
     1381 ggttcaggcc ttggaggagt tgctgccatc cccgaacttc ggggtgaagc tggagacgct
     1441 gcccctgtcc cctggcagcg gggccgagcc cgacaccaag ccggtcttcc atactctcac
     1501 caacggccac gccaagggcc gcttgtccgg cgacgagggc tccgaggacg gcgacgacta
     1561 tgacacacct cccatcctca aggagctgca ggcgctcaac accgaggagg cggcggagca
     1621 gcgggcggag gtggaccgga tgctcagtga ggacccttgg agggctgcta aaatgatcaa
     1681 gggttacatg cagcaacaca acatccccca gagggaggtg gtcgatgtca ccggcctgaa
     1741 ccagtcgcac ctctcccagc atctcaacaa gggcacccct atgaagaccc agaagcgtgc
     1801 cgctctgtac acctggtacg tcagaaagca acgagagatc ctccgacaat tcaaccagac
     1861 agtccagagt tctggaaata tgacagacaa aagcagtcag gatcagctgc tgtttctctt
     1921 tccagagttc agtcaacaga gccatgggcc tgggcagtcc gatgatgcct gctctgagcc
     1981 caccaacaag aagatgcgcc gcaaccggtt caaatggggg cccgcgtccc agcaaatctt
     2041 gtaccaggcc tacgatcggc aaaagaaccc cagcaaggaa gagagagagg ggccttagtg
     2101 gaggaatgca acagggcaga atgtttgcag cgaggggtgt ccccctccaa agcccacggc
     2161 ctgggctcca acttggtcac tgaggtccgt gtctacaact ggtttgcaaa ccgcaggaag
     2221 gaggaggcat tccggcaaaa gctggccatg gacgcctata gctccaacca gactcacagc
     2281 ctgaaccctc tgctctccca cggctccccc caccaccagc ccagctcctc tcctccaaac
     2341 aagctgtcag gagtgcgcta cagccagcag ggaaacaatg agatcacttc ctcctcaaca
     2401 atcagtcacc atggcaacag cgccatggtg accagccagt cggttttaca gcaagtctcc
     2461 ccagccagcc tggacccagg ccacaatctc ctctcacctg atggtaaaat gatctcagtc
     2521 tcaggaggag gtttgccccc agtcagcacc ttgacgaata tccacagcct ctcccaccat
     2581 aatccccagc aatctcaaaa cctcatcatg acacccctct ctggagtcat ggcaattgca
     2641 caaagcctca acacctccca agcacagagt gtccctgtca tcaacagtgt ggccggcagc
     2701 ctggcagccc tgcagcccgt ccagttctcc cagcagctgc acagccctca ccagcagccc
     2761 ctcatgcagc agagcccagg cagccacatg gcccagcagc ccttcatggc agctgtgact
     2821 cagctgcaga actcacacat gtacgcacac aagcaggaac ccccccagta ttcccacacc
     2881 tcccggtttc catctgcaat ggtggtcaca gataccagca gcatcagtac actcaccaac
     2941 atgtcttcaa gtaaacagtg tcctctacaa gcctggtgat gcccacacac cacttacttc
     3001 gtgcgcaaca acaaggaccc tgttttccac accatcaccc tctgggcagc tgtcatggaa
     3061 aagcccagtg acctgaccag cacctgcgag aggtccctgc ttacctgacg gacgtctaga
     3121 gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     3181 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3241 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3301 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3361 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3421 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3481 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3541 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3601 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3661 tgatggttca cgtacctaga agttcctatt ccgaagttcc tattctctag aaagtatagg
     3721 aacttccttg gccaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     3781 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     3841 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     3901 ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt
     3961 gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     4021 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     4081 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     4141 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     4201 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     4261 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     4321 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     4381 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     4441 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     4501 aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     4561 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     4621 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg
     4681 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     4741 gagggcaaag gaatagcacg tactacgaga tttcgattcc accgccgcct tctatgaaag
     4801 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4861 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4921 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4981 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5041 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5101 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5161 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5221 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5281 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5341 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5401 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5461 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5521 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5581 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5641 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5701 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5761 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5821 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5881 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5941 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6001 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6061 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6121 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6181 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6241 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6301 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6361 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6421 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6481 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6541 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6601 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6661 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6721 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6781 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6841 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6901 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6961 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7021 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7081 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7141 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7201 gccacctgac gtc
//
