LOCUS       sequence-14284-e        7413 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 7413)
  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 7413)
  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_HNF4A8
FEATURES             Location/Qualifiers
     source          1..7413
                     /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(3365..3382)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3371..3595
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3728..3747)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3879..3926
                     /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             3934..4956
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3934..4956
                     /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    5086..5207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5123..5142)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5177..5196
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5256..5272)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5256..5272)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5269..5291)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5280..5296
                     /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(5304..5310,5311..5328,5329..5334))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5349..5370
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5487..5504)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5658..6246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5738..5757)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6417..7208,7209..7277))
                     /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     7040..7059
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7278..7382)
                     /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 gtaccatggt cagcgtgaac gcgcccctcg gggctccagt ggagagttct tacgacacgt
     1381 ccccatcaga aggcaccaac ctcaacgcgc ccaacagcct gggtgtcagc gccctgtgtg
     1441 ccatctgcgg ggaccgggcc acgggcaaac actacggtgc ctcgagctgt gacggctgca
     1501 agggcttctt ccggaggagc gtgcggaaga accacatgta ctcctgcaga tttagccggc
     1561 agtgcgtggt ggacaaagac aagaggaacc agtgccgcta ctgcaggctc aagaaatgct
     1621 tccgggctgg catgaagaag gaagccgtcc agaatgagcg ggaccggatc agcactcgaa
     1681 ggtcaagcta tgaggacagc agcctgccct ccatcaatgc gctcctgcag gcggaggtcc
     1741 tgtcccgaca gatcacctcc cccgtctccg ggatcaacgg cgacattcgg gcgaagaaga
     1801 ttgccagcat cgcagatgtg tgtgagtcca tgaaggagca gctgctggtt ctcgttgagt
     1861 gggccaagta catcccagct ttctgcgagc tccccctgga cgaccaggtg gccctgctca
     1921 gagcccatgc tggcgagcac ctgctgctcg gagccaccaa gagatccatg gtgttcaagg
     1981 acgtgctgct cctaggcaat gactacattg tccctcggca ctgcccggag ctggcggaga
     2041 tgagccgggt gtccatacgc atccttgacg agctggtgct gcccttccag gagctgcaga
     2101 tcgatgacaa tgagtatgcc tacctcaaag ccatcatctt ctttgaccca gatgccaagg
     2161 ggctgagcga tccagggaag atcaagcggc tgcgttccca ggtgcaggtg agcttggagg
     2221 actacatcaa cgaccgccag tatgactcgc gtggccgctt tggagagctg ctgctgctgc
     2281 tgcccacctt gcagagcatc acgtggcaga tgatcgagca gatccagttc atcaagctct
     2341 tcggcatggc caagattgac aacctgttgc aggagatgct gctgggaggg tcccccagcg
     2401 atgcacccca tgcccaccac cccctgcacc ctcacctgat gcaggaacat atgggaacca
     2461 acgtcatcgt tgccaacaca atgcccactc acctcagcaa cggacagatg tgtgagtggc
     2521 cccgacccag gggacaggca gccacccctg agaccccaca gccctcaccg ccaggtggct
     2581 cagggtctga gccctataag ctcctgccgg gagccgtcgc cacaatcgtc aagcccctct
     2641 ctgccatccc ccagccgacc atcaccaagc aggaagttat ctagcaagcc gctggggctt
     2701 gggggctcca ctggctcccc ccagccccct aagagagcac ctggtgatca cgtggtcacg
     2761 gcaaaggaag acgtgatgcc aggaccagtc ccagagcagg aatgggaagg atgaagggcc
     2821 cgagaacatg gcctaagggc cacatcccac tgccaccctt gacgccctgc tctggataac
     2881 aagactttga cttggggaga cctctactgc cttggacaac ttttctcatg ttgaagccac
     2941 tgccttcacc ttcaccttca tccatgtcca acccccgact tcatcccaaa ggacagccgc
     3001 ctggagatga cttgaggcct tacttaaacc cagctccctt cttccctagc ctggtgcttc
     3061 tcctctccta gccccggtca tggtgtccag acagagccct gtgaggctgg gtccaattgt
     3121 ggcacttggg gcaccttgct cctccttctg ctgctgcccc cacctctgct gcctccctct
     3181 gctgtcacct tgctcagcca tcccgtcttc tccaacacca cctctacaga ggccaaggag
     3241 gccttggaaa cgattccccc agtcattctg ggaacatgtt gtaagcactg actgggacca
     3301 ggcaccaggc agggtctaga gcggccgctc gagtctagag ggcccgttta aacccgctga
     3361 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3421 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3481 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3541 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3601 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3661 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3721 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3781 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3841 cccaaaaaac ttgattaggg tgatggttca cgtacctaga agttcctatt ccgaagttcc
     3901 tattctctag aaagtatagg aacttccttg gccaaaaagc ctgaactcac cgcgacgtct
     3961 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag
     4021 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
     4081 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc
     4141 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc
     4201 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
     4261 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg
     4321 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc
     4381 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc
     4441 agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa
     4501 gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc
     4561 ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc
     4621 aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag
     4681 cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt
     4741 cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag
     4801 ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc
     4861 cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac
     4921 cgacgcccca gcactcgtcc gagggcaaag gaatagcacg tactacgaga tttcgattcc
     4981 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5041 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5101 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5161 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5221 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5281 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5341 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5401 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5461 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5521 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5581 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5641 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5701 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5761 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5821 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5881 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5941 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6001 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6061 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6121 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6181 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6241 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6301 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6361 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6421 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6481 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6541 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6601 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6661 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6721 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6781 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6841 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6901 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6961 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7021 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7081 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7141 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7201 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7261 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7321 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7381 cgcacatttc cccgaaaagt gccacctgac gtc
//
