LOCUS       sequence-14285-e        7479 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 7479)
  AUTHORS   Erdmann S, Senkel S, Arndt T, Lucas B, Lausen J, Klein-Hitpass L, 
            Ryffel GU, Thomas H
  TITLE     Tissue-specific transcription factor HNF4alpha inhibits cell 
            proliferation and induces apoptosis in the pancreatic INS-1 
            beta-cell line.
  JOURNAL   Biol Chem. 2007 Jan . 388(1):91-106.
  PUBMED    17214554
REFERENCE   2  (bases 1 to 7479)
  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: "Biol Chem. 
            2007 Jan . 388(1):91-106."
COMMENT     Sequence Label: FR_HNF4A2-C106R
FEATURES             Location/Qualifiers
     source          1..7479
                     /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             1059..1088
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1098..1127
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1137..1166
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1176..1205
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1215..1244
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1278..1307
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3431..3448)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3437..3661
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3794..3813)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3945..3992
                     /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             4000..5022
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4000..5022
                     /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    5152..5273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5189..5208)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5243..5262
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5322..5338)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5322..5338)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5335..5357)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5346..5362
                     /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(5370..5376,5377..5394,5395..5400))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5415..5436
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5553..5570)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5724..6312)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5804..5823)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6483..7274,7275..7343))
                     /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     7106..7125
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7344..7448)
                     /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 cactagtcca gtgtggtgga
     1021 attctgcaga tatccagcac agtggcggcc gcgctatgga gcaaaagctc atttctgaag
     1081 aggacttgaa tgaaatggag caaaagctca tttctgaaga ggacttgaat gaaatggagc
     1141 aaaagctcat ttctgaagag gacttgaatg aaatggagca aaagctcatt tctgaagagg
     1201 acttgaatga aatggagcaa aagctcattt ctgaagagga cttgaatgaa atggagagct
     1261 tgggcgacct caccatggag caaaagctca tttctgaaga ggacttgaat tcaaggcctg
     1321 gtaccatgcg actctccaaa accctcgtcg acatggacat ggccgactac agtgctgcac
     1381 tggacccagc ctacaccacc ctggaatttg agaatgtgca ggtgttgacg atgggcaatg
     1441 acacgtcccc atcagaaggc accaacctca acgcgcccaa cagcctgggt gtcagcgccc
     1501 tgtgtgccat ctgcggggac cgggccacgg gcaaacacta cggtgcctcg agctgtgacg
     1561 gctgcaaggg cttcttccgg aggagcgtgc ggaagaacca catgtactcc tgcagattta
     1621 gccggcagtg cgtggtggac aaagacaaga ggaaccagtg ccgctaccgc aggctcaaga
     1681 aatgcttccg ggctggcatg aagaaggaag ccgtccagaa tgagcgggac cggatcagca
     1741 ctcgaaggtc aagctatgag gacagcagcc tgccctccat caatgcgctc ctgcaggcgg
     1801 aggtcctgtc ccgacagatc acctcccccg tctccgggat caacggcgac attcgggcga
     1861 agaagattgc cagcatcgca gatgtgtgtg agtccatgaa ggagcagctg ctggttctcg
     1921 ttgagtgggc caagtacatc ccagctttct gcgagctccc cctggacgac caggtggccc
     1981 tgctcagagc ccatgctggc gagcacctgc tgctcggagc caccaagaga tccatggtgt
     2041 tcaaggacgt gctgctccta ggcaatgact acattgtccc tcggcactgc ccggagctgg
     2101 cggagatgag ccgggtgtcc atacgcatcc ttgacgagct ggtgctgccc ttccaggagc
     2161 tgcagatcga tgacaatgag tatgcctacc tcaaagccat catcttcttt gacccagatg
     2221 ccaaggggct gagcgatcca gggaagatca agcggctgcg ttcccaggtg caggtgagct
     2281 tggaggacta catcaacgac cgccagtatg actcgcgtgg ccgctttgga gagctgctgc
     2341 tgctgctgcc caccttgcag agcatcacgt ggcagatgat cgagcagatc cagttcatca
     2401 agctcttcgg catggccaag attgacaacc tgttgcagga gatgctgctg ggagggtccc
     2461 ccagcgatgc accccatgcc caccaccccc tgcaccctca cctgatgcag gaacatatgg
     2521 gaaccaacgt catcgttgcc aacacaatgc ccactcacct cagcaacgga cagatgtgtg
     2581 agtggccccg acccagggga caggcagcca cccctgagac cccacagccc tcaccgccag
     2641 gtggctcagg gtctgagccc tataagctcc tgccgggagc cgtcgccaca atcgtcaagc
     2701 ccctctctgc catcccccag ccgaccatca ccaagcagga agttatctag caagccgctg
     2761 gggcttgggg gctccactgg ctccccccag ccccctaaga gagcacctgg tgatcacgtg
     2821 gtcacggcaa aggaagacgt gatgccagga ccagtcccag agcaggaatg ggaaggatga
     2881 agggcccgag aacatggcct aagggccaca tcccactgcc acccttgacg ccctgctctg
     2941 gataacaaga ctttgacttg gggagacctc tactgccttg gacaactttt ctcatgttga
     3001 agccactgcc ttcaccttca ccttcatcca tgtccaaccc ccgacttcat cccaaaggac
     3061 agccgcctgg agatgacttg aggccttact taaacccagc tcccttcttc cctagcctgg
     3121 tgcttctcct ctcctagccc cggtcatggt gtccagacag agccctgtga ggctgggtcc
     3181 aattgtggca cttggggcac cttgctcctc cttctgctgc tgcccccacc tctgctgcct
     3241 ccctctgctg tcaccttgct cagccatccc gtcttctcca acaccacctc tacagaggcc
     3301 aaggaggcct tggaaacgat tcccccagtc attctgggaa catgttgtaa gcactgactg
     3361 ggaccaggca ccaggcaggg tctagagcgg ccgctcgagt ctagagggcc cgtttaaacc
     3421 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3481 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3541 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3601 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3661 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3721 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3781 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3841 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3901 ctcgacccca aaaaacttga ttagggtgat ggttcacgta cctagaagtt cctattccga
     3961 agttcctatt ctctagaaag tataggaact tccttggcca aaaagcctga actcaccgcg
     4021 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc
     4081 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg
     4141 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca
     4201 tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc
     4261 tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg
     4321 cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc
     4381 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt
     4441 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac
     4501 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc
     4561 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat
     4621 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag
     4681 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac
     4741 ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc
     4801 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg
     4861 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa
     4921 atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt
     4981 ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agcacgtact acgagatttc
     5041 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5101 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5161 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5221 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5281 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5341 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5401 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5461 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5521 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5581 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5641 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5701 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5761 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5821 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5881 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5941 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6001 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6061 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6121 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6181 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6241 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6301 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6361 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6421 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6481 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6541 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6601 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6661 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6721 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6781 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6841 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6901 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6961 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7021 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7081 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7141 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7201 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7261 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7321 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7381 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7441 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
