LOCUS       sequence-10161-e        8890 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 8890)
  AUTHORS   Tussie-Luna MI, Bayarsaihan D, Ruddle FH, Roy AL
  TITLE     Repression of TFII-I-dependent transcription by nuclear exclusion.
  JOURNAL   Proc Natl Acad Sci U S A. 2001 Jul 3. 98(14):7789-94.
  PUBMED    11438732
REFERENCE   2  (bases 1 to 8890)
  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: "Proc Natl 
            Acad Sci U S A. 2001 Jul 3. 98(14):7789-94."
COMMENT     Sequence Label: pEBG-hBEN
FEATURES             Location/Qualifiers
     source          1..8890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        252..462
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        274..470
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      321..456
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        557..1739
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          788..1730
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1687..1707
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1791..2444
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MAPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2402..2424
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2451..2468
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5322..5339
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6024..6041)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6024..6040)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6033..6055)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6253..6708
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6340..6359)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6550..6571
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6725..6744)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6844..6866
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6904..6922)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6990..7094
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7095..7163,7164..7955)
                     /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     complement(7313..7332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8126..8714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8615..8634
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8868..8885
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttc
      241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
      301 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
      361 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
      421 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctttgca
      481 aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt
      541 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
      601 ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg
      661 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
      721 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
      781 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
      841 ttgcgtgcct tgaattactt ccacgcccct ggctgcagta cgtgattctt gatcccgagc
      901 ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc
      961 gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc
     1021 ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg
     1081 ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg
     1141 gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt
     1201 cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg
     1261 gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa
     1321 ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg
     1381 cagggagctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac
     1441 aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg
     1501 cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg
     1561 gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc
     1621 cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt
     1681 tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag
     1741 gaattctcta gagatccctc gacctcgaga tccattgtgc tggatctaca atggccccta
     1801 tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc
     1861 ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca
     1921 aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta
     1981 aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg
     2041 gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat
     2101 acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta
     2161 gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa
     2221 atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat
     2281 acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg
     2341 aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc
     2401 agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggttccgc
     2461 gtggatccac tagtggtacc atcgatggaa tggccttgct gggtaagcgc tgtgacgtcc
     2521 ccaccaacgg ctgcggaccc gaccgctgga actccgcgtt cacccgcaaa gacgagatca
     2581 tcaccagcct cgtgtctgcc ttagactcca tgtgctcagc gctgtccaaa ctgaacgccg
     2641 aggtggcctg tgtcgccgtg cacgatgaga gcgcctttgt ggtgggcaca gagaagggga
     2701 gaatgttcct gaatgcccgg aaggagctac agtcagactt cctcaggttc tgccgagggc
     2761 ccccgtggaa ggatccggag gcagagcacc ccaagaaggt gcagcggggc gagggtggaa
     2821 gccgtagcct ccctcggtcc tccctggaac atggctcaga tgtgtacctt ctgcggaaga
     2881 tggtagagga ggtgtttgat gttctttata gcgaggccct gggaagggcc agtgtggtgc
     2941 cactgcccta tgagaggctg ctcagggagc cagggctgct ggccgtgcag gggctgcccg
     3001 aaggcctggc cttccgaagg ccagccgagt atgaccccaa ggccctcatg gccatcctgg
     3061 aacacagcca ccgcatccgc ttcaagctca agaggccact tgaggatggc gggcgggact
     3121 cgaaggccct ggtggagctg aacggtgtct ccctgattcc caaggggtca cgggactgtg
     3181 gcctgcatgg ccaggccccc aaggtgccac cccaggacct gcccccaacc gccacctcct
     3241 cctccatggc cagcttcctg tacagcacgg cgctccccaa ccacgccatc cgagagctca
     3301 agcaggaagc accttcctgc ccccttgccc ccagcgacct gggcctgagt cggcccatgc
     3361 cagagcccaa ggccaccggt gcccaagact tctccgactg ttgtggacag aagcccactg
     3421 ggcctggtgg gcctctcatc cagaacgtcc atgcctccaa gcgcattctc ttctccatcg
     3481 tccatgacaa gtcagagaag tgggacgcct tcataaagga aaccgaggac atcaacacgc
     3541 tccgggagtg tgtgcagatc ctgtttaaca gcagatatgc ggaagccctg ggcctggacc
     3601 acatggtccc cgtgccctac cggaagattg cctgtgaccc ggaggctgtg gagatcgtgg
     3661 gcatcccgga caagatcccc ttcaagcgcc cctgcactta tggagtcccc aagctgaagc
     3721 ggatcctgga ggagcgccat agtatccact tcatcattaa gaggatgttt gatgagcgaa
     3781 ttttcacagg gaacaagttt accaaagaca ccacgaagct ggagccagcc agcccgccag
     3841 aggacacctc tgcagaggtc tctagggcca ccgtccttga ccttgctggg aatgctcggt
     3901 cagacaaggg cagcatgtct gaagactgtg ggccaggaac gtccggggag ctgggcgggc
     3961 tgaggccgat caaaattgag ccagaggatc tggacatcat tcaggtcacc gtcccagacc
     4021 cctcgccaac ctctgaggaa atgacagact cgatgcctgg gcacctgcca tcggaggatt
     4081 ctggttatgg gatggagatg ctgacagaca aaggtctgag tgaggacgcg cggccggagg
     4141 agaggcccgt ggaggacagc cacggtgacg tgatccggcc cctgcggaag caggtggagc
     4201 tgctcttcaa cacacgatac gccaaggcca ttggcatctc ggagcccgtc aaggtgccgt
     4261 actccaagtt tctgatgcac ccggaggagc tgtttgtggt gggactgcct gaaggcatct
     4321 ccctccgcag gcccaactgc ttcgggatcg ccaagctccg gaagattctg gaggccagca
     4381 acagcatcca gtttgtcatc aagaggcccg agctgctcac tgagggagtc aaagagccca
     4441 tcatggatag tcaagagagg gattccgggg accctctggt ggacgagagc ctgaagagac
     4501 agggctttca agaaaattat gacgcgaggc tctcacggat cgacatcgcc aacacactaa
     4561 gggagcaggt ccaggacctt ttcaataaga aatacgggga agccttgggc atcaagtacc
     4621 cggtccaggt cccctacaag cggatcaaga gtaaccccgg ctccgtgatc atcgaggggc
     4681 tgcccccagg aatcccgttc cgaaagccct gtaccttcgg ctcccagaac ctggagagga
     4741 ttcttgctgt ggctgacaag atcaagttca cagtcaccag gcctttccaa ggactcatcc
     4801 caaagcctga tgaagatgac gccaacagac tcggggagaa ggtgatcctg cgggagcagg
     4861 tgaaggaact cttcaacgag aaatacggtg aggccctggg cctgaaccgg ccggtgctgg
     4921 tcccttataa actaatccgg gacagcccag acgccgtgga ggtcacgggt ctgcctgatg
     4981 acatcccctt ccggaacccc aacacgtacg acatccaccg gctggagaag atcctgaagg
     5041 cccgagagca tgtccgcatg gtcatcatta accagctcca accctttgca gaaatctgca
     5101 atgatgccaa ggtgccagcc aaagacagca gcattcccaa gcgcaagaga aagcgggtct
     5161 cggaaggaaa ttccgtctcc tcttcctcct cgtcttcctc ttcctcgtcc tctaacccgg
     5221 attcagtggc atcggccaac cagatctcac tcgtgcaatg gccaatgtac atggtggact
     5281 atgccggcct gaacgtgcag ctcccgggac ctcttaatta ccaccaccac caccaccact
     5341 aggcggccgc gactctagag tgagggtccc cacctgggac ccttgagagt atcaggtctc
     5401 ccacgtggga gacaagaaat ccctgtttaa tatttaaaca gcagtgttcc ccatctgggt
     5461 ccttgcaccc ctcactctgg cctcagccga ctgcacagcg gcccctgcat ccccttggct
     5521 gtgaggcccc tggacaagca gaggtggcca gagctgggag gcatggccct ggggtcccac
     5581 gaatttgctg gggaatctcg tttttcttct taagactttt gggacatggt ttgactcccg
     5641 aacatcaccg acgcgtctcc tgtttttctg ggtggcctcg ggacacctgc cctgccccca
     5701 cgagggtcag gactgtgact ctttttaggg ccaggcaggt gcctggacat ttgccttgct
     5761 ggacggggac tggggatgtg ggagggagca gacaggagga atcatgtcag gcctgtgtgt
     5821 gaaaggaagc tccactgtca ccctccacct cttcaccccc cactcaccag tgtcccctcc
     5881 actgtcacat tgtaactgaa cttcaggata ataaagtgtt tgcctccaaa aaaaaaaaaa
     5941 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6001 aaaaaaaaaa aaaaaaagaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     6061 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     6121 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     6181 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     6241 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     6301 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     6361 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     6421 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     6481 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     6541 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt
     6601 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6661 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     6721 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     6781 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     6841 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     6901 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     6961 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     7021 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     7081 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     7141 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     7201 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     7261 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     7321 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     7381 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     7441 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     7501 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     7561 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     7621 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     7681 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     7741 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     7801 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     7861 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7921 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     7981 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     8041 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     8101 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     8161 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     8221 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     8281 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     8341 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     8401 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     8461 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     8521 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     8581 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     8641 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     8701 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     8761 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     8821 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     8881 gcgaggaagc
//
