LOCUS       sequence-14273-e        7211 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 7211)
  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 7211)
  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
FEATURES             Location/Qualifiers
     source          1..7211
                     /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(3163..3180)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3169..3393
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3526..3545)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3677..3724
                     /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             3732..4754
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3732..4754
                     /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    4884..5005
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4921..4940)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4975..4994
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5054..5070)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5054..5070)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5067..5089)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5078..5094
                     /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(5102..5108,5109..5126,5127..5132))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5147..5168
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5285..5302)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5456..6044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5536..5555)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6215..7006,7007..7075))
                     /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     6838..6857
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7076..7180)
                     /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 ccttagtgga
     2101 ggaatgcaac agggcagaat gtttgcagcg aggggtgtcc ccctccaaag cccacggcct
     2161 gggctccaac ttggtcactg aggtccgtgt ctacaactgg tttgcaaacc gcaggaagga
     2221 ggaggcattc cggcaaaagc tggccatgga cgcctatagc tccaaccaga ctcacagcct
     2281 gaaccctctg ctctcccacg gctcccccca ccaccagccc agctcctctc ctccaaacaa
     2341 gctgtcagga gtgcgctaca gccagcaggg aaacaatgag atcacttcct cctcaacaat
     2401 cagtcaccat ggcaacagcg ccatggtgac cagccagtcg gttttacagc aagtctcccc
     2461 agccagcctg gacccaggcc acaatctcct ctcacctgat ggtaaaatga tctcagtctc
     2521 aggaggaggt ttgcccccag tcagcacctt gacgaatatc cacagcctct cccaccataa
     2581 tccccagcaa tctcaaaacc tcatcatgac acccctctct ggagtcatgg caattgcaca
     2641 aagcctcaac acctcccaag cacagagtgt ccctgtcatc aacagtgtgg ccggcagcct
     2701 ggcagccctg cagcccgtcc agttctccca gcagctgcac agccctcacc agcagcccct
     2761 catgcagcag agcccaggca gccacatggc ccagcagccc ttcatggcag ctgtgactca
     2821 gctgcagaac tcacacatgt acgcacacaa gcaggaaccc ccccagtatt cccacacctc
     2881 ccggtttcca tctgcaatgg tggtcacaga taccagcagc atcagtacac tcaccaacat
     2941 gtcttcaagt aaacagtgtc ctctacaagc ctggtgatgc ccacacacca cttacttcgt
     3001 gcgcaacaac aaggaccctg ttttccacac catcaccctc tgggcagctg tcatggaaaa
     3061 gcccagtgac ctgaccagca cctgcgagag gtccctgctt acctgacgga cgtctagagc
     3121 ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     3181 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3241 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3301 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3361 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3421 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3481 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3541 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3601 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3661 atggttcacg tacctagaag ttcctattcc gaagttccta ttctctagaa agtataggaa
     3721 cttccttggc caaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
     3781 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
     3841 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
     3901 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
     3961 ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
     4021 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
     4081 ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac
     4141 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
     4201 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
     4261 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     4321 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     4381 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     4441 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     4501 gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     4561 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     4621 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga
     4681 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     4741 gggcaaagga atagcacgta ctacgagatt tcgattccac cgccgccttc tatgaaaggt
     4801 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4861 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4921 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4981 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5041 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5101 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5161 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5221 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5281 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5341 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5401 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5461 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5521 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5581 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5641 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5701 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5761 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5821 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5881 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5941 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6001 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6061 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6121 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6181 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6241 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6301 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6361 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6421 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6481 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6541 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6601 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6661 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6721 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6781 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6841 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6901 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6961 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7021 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7081 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7141 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7201 cacctgacgt c
//
