LOCUS       sequence-14271-e        6629 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 6629)
  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 6629)
  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_HNF6
FEATURES             Location/Qualifiers
     source          1..6629
                     /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             1004..1021
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1409..1438
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1412..1441
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(2581..2598)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2587..2811
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2944..2963)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3095..3142
                     /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             3150..4172
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3150..4172
                     /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    4302..4423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4339..4358)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4393..4412
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4472..4488)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4472..4488)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4485..4507)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4496..4512
                     /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(4520..4526,4527..4544,4545..4550))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4565..4586
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4703..4720)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4874..5462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4954..4973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5633..6424,6425..6493))
                     /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     6256..6275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6494..6598)
                     /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 ctatcgatac catgcggggt tctcatcatc atcatcatca
     1021 tggtgaagct ttgatagaca tgaacgcgca gctgaccatg gaagcgatcg gcgagctgca
     1081 cggggtgagc catgagccgg tgcccgcccc tgccgacctg ctgggcggca gcccccacgc
     1141 gcgcagctcc gtggcgcacc gcggcagcca cctgcccccc gcgcacccgc gctccatggg
     1201 catggcgtcc ctgctggacg gcggcagcgg cggcggagat taccaccacc accaccgggc
     1261 ccctgagcac agcctggccg gccccctgca tcccaccatg accatggcct gcgagactcc
     1321 cccaggtatg agcatgccca ccacctacac caccttgacc cctctgcagc cgctgcctcc
     1381 catctccaca gtctcggaca agttccccca ccatcaccac caccaccatc accaccacca
     1441 cccgcaccac caccagcgcc tggcgggcaa cgtgagcggt agcttcacgc tcatgcggga
     1501 tgagcgcggg ctggcctcca tgaataacct ctataccccc taccacaagg acgtggccgg
     1561 catgggccag agcctctcgc ccctctccag ctccggtctg ggcagcatcc acaactccca
     1621 gcaagggctc ccccactatg cccacccggg ggccgccatg cccaccgaca agatgctcac
     1681 ccccaacggc ttcgaagccc accacccggc catgctcggc cgccacgggg agcagcacct
     1741 cacgcccacc tcggccggca tggtgcccat caacggcctt cctccgcacc atccccacgc
     1801 ccacctgaac gcccagggcc acgggcaact cctgggcaca gcccgggagc ccaacccttc
     1861 ggtgaccggc gcgcaggtca gcaatggaag taattcaggg cagatggaag agatcaatac
     1921 caaagaggtg gcgcagcgta tcaccaccga gctcaagcgc tacagcatcc cacaggccat
     1981 cttcgcgcag agggtgctct gccgctccca ggggaccctc tcggacctgc tgcgcaaccc
     2041 caaaccctgg agcaaactca aatccggccg ggagaccttc cggaggatgt ggaagtggct
     2101 gcaggagccg gagttccagc gcatgtccgc gctccgctta gcagcatgca aaaggaaaga
     2161 acaagaacat gggaaggata gaggcaacac acccaaaaag cccaggttgg tcttcacaga
     2221 tgtccagcgt cgaactctac atgcaatatt caaggaaaat aagcgtccat ccaaagaatt
     2281 gcaaatcacc atttcccagc agctggggtt ggagctgagc actgtcagca acttcttcat
     2341 gaacgcaaga aggaggagtc tggacaagtg gcaggacgag ggcagctcca attcaggcaa
     2401 ctcatcttct tcatcaagca cttgtaccaa agcatgaagg aagaaccaca aactaaaacc
     2461 tcggtggaaa agcttggtac cgagctcgga tccactagtc cagtgtggtg gaattctgca
     2521 gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag
     2581 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2641 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2701 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2761 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     2821 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     2881 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2941 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3001 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3061 aaaaacttga ttagggtgat ggttcacgta cctagaagtt cctattccga agttcctatt
     3121 ctctagaaag tataggaact tccttggcca aaaagcctga actcaccgcg acgtctgtcg
     3181 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     3241 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     3301 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     3361 tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct
     3421 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     3481 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     3541 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     3601 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     3661 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     3721 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     3781 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     3841 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     3901 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     3961 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     4021 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     4081 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     4141 gccccagcac tcgtccgagg gcaaaggaat agcacgtact acgagatttc gattccaccg
     4201 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4261 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4321 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4381 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4441 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4501 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4561 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4621 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4681 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4741 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4801 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4861 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4921 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4981 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5041 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5101 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5161 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5221 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5281 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5341 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5401 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5461 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5521 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5581 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5641 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5701 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5761 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5821 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5881 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5941 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6001 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6061 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6121 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6181 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6241 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6301 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6361 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6421 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6481 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6541 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6601 catttccccg aaaagtgcca cctgacgtc
//
