LOCUS       sequence-163059-        7495 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 7495)
  AUTHORS   Senkel S, Lucas B, Klein-Hitpass L, Ryffel GU
  TITLE     Identification of target genes of the transcription factor HNF1beta 
            and HNF1alpha in a human embryonic kidney cell line.
  JOURNAL   Biochim Biophys Acta. 2005 Dec 20. 1731(3):179-90.
  PUBMED    16297991
REFERENCE   2  (bases 1 to 7495)
  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: "Biochim 
            Biophys Acta. 2005 Dec 20. 1731(3):179-90."
COMMENT     Sequence Label: FR_HNF1A
FEATURES             Location/Qualifiers
     source          1..7495
                     /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"
     regulatory      2774..2783
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3447..3464)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3453..3677
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3810..3829)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3961..4008
                     /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             4016..5038
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4016..5038
                     /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    5168..5289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5205..5224)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5259..5278
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5338..5354)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5338..5354)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5351..5373)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5362..5378
                     /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(5386..5392,5393..5410,5411..5416))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5431..5452
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5569..5586)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5740..6328)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5820..5839)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6499..7290,7291..7359))
                     /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     7122..7141
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7360..7464)
                     /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 tccatggttt ctaaactgag
     1321 ccagctgcag acggagctcc tggcggccct gctggagtca gggctgagca aagaggcact
     1381 gctccaggca ctgggtgagc cggggcccta cctcctggct ggagaaggcc ccctggacaa
     1441 gggggagtcc tgcggcggcg gtcgagggga gctggctgag ctgcccaatg ggctggggga
     1501 gactcggggc tccgaggacg agacggacga cgatggggaa gacttcacgc cacccatcct
     1561 caaagagctg gagaacctca gccctgagga ggcggcccac cagaaagccg tggtggagac
     1621 ccttctgcag gaggacccgt ggcgtgtggc gaagatggtc aagtcctacc tgcagcagca
     1681 caacatccca cagcgggagg tggtcgatac cactggcctc aaccagtccc acctgtccca
     1741 acacctcaac aagggcactc ccatgaagac gcagaagcgg gccgccctgt acacctggta
     1801 cgtccgcaag cagcgagagg tggcgcagca gttcacccat gcagggcagg gagggctgat
     1861 tgaagagccc acaggtgatg agctaccaac caagaagggg cggaggaacc gtttcaagtg
     1921 gggcccagca tcccagcaga tcctgttcca ggcctatgag aggcagaaga accctagcaa
     1981 ggaggagcga gagacgctag tggaggagtg caatagggcg gaatgcatcc agagaggggt
     2041 gtccccatca caggcacagg ggctgggctc caacctcgtc acggaggtgc gtgtctacaa
     2101 ctggtttgcc aaccggcgca aagaagaagc cttccggcac aagctggcca tggacacgta
     2161 cagcgggccc cccccagggc caggcccggg acctgcgctg cccgctcaca gctcccctgg
     2221 cctgcctcca cctgccctct cccccagtaa ggtccacggt gtgcgctttg gacagcctgc
     2281 gaccagtgag actgcagaag taccctcaag cagcggcggt cccttagtga cagtgtctac
     2341 acccctccac caagtgtccc ccacgggcct ggagcccagc cacagcctgc tgagtacaga
     2401 agccaagctg gtctcagcag ctgggggccc cctcccccct gtcagcaccc tgacagcact
     2461 gcacagcttg gagcagacat ccccaggcct caaccagcag ccccagaacc tcatcatggc
     2521 ctcacttcct ggggtcatga ccatcgggcc tggtgagcct gcctccctgg gtcctacgtt
     2581 caccaacaca ggtgcctcca ccctggtcat cggcctggcc tccacgcagg cacagagtgt
     2641 gccggtcatc aacagcatgg gcagcagcct gaccaccttg cagcccgtcc agttctccca
     2701 gccgctgcac ccctcctacc agcagccgct catgccacct gtgcagagcc atgtgaccca
     2761 gaaccccttc atggccacca tggctcagct gcagagcccc cacgccctct acagccacaa
     2821 gcccgaggtg gcccagtaca cccacacggg cctgctcccg cagactatgc tcatcaccga
     2881 caccaccaac ctgagcgccc tggccagcct cacgcccacc aagcaggtct tcacctcaga
     2941 cactgaggcc tccagtgagt ccgggcttca cacgccggca tctcaggcca ccaccctcca
     3001 cgtccccagc caggaccctg ccggcatcca gcacctgcag ccggcccacc ggctcagcgc
     3061 cagccccaca gtgtcctcca gcagcctggt gctgtaccag agctcagact ccagcaatgg
     3121 ccagagccac ctgctgccat ccaaccacag cgtcatcgag accttcatct ccacccagat
     3181 ggcctcttcc tcccagtaac cacggcacct gggccctggg gcctgtactg cctgcttggg
     3241 gggtgatgag ggcagcagcc agccctgcct ggaggacctg agcctgccga gcaaccgtgg
     3301 cccttcctgg acagctgtgc ctcgctcccc actctgctct gatgcagccc gggggatcca
     3361 ctagtccagt gtggtggaat tctgcagata tccagcacag tggcggccgc tcgagtctag
     3421 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3481 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3541 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3601 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3661 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3721 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3781 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3841 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3901 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaccta
     3961 gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcct tggccaaaaa
     4021 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     4081 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     4141 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     4201 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt
     4261 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     4321 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
     4381 tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     4441 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     4501 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     4561 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     4621 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     4681 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     4741 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     4801 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     4861 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     4921 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     4981 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca
     5041 cgtactacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5101 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5161 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5221 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5281 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5341 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5401 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5461 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5521 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5581 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5641 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5701 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5761 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5821 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5881 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5941 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6001 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6061 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6121 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6181 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6241 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6301 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6361 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6421 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6481 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6541 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6601 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6661 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6721 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6781 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6841 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6901 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6961 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7021 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7081 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7141 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7201 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7261 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7321 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7381 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7441 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
