LOCUS       sequence-10165-e        9421 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 9421)
  AUTHORS   Cheriyath V, Novina CD, Roy AL
  TITLE     TFII-I regulates Vbeta promoter activity through an initiator 
            element.
  JOURNAL   Mol Cell Biol. 1998 Aug . 18(8):4444-54.
  PUBMED    9671454
REFERENCE   2  (bases 1 to 9421)
  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: "Mol Cell 
            Biol. 1998 Aug . 18(8):4444-54."
COMMENT     Sequence Label: pEBG-TFII-I delta isoform
FEATURES             Location/Qualifiers
     source          1..9421
                     /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             2484..2501
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2511..2528
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(6555..6572)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6555..6571)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6564..6586)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6784..7239
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6871..6890)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7081..7102
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7256..7275)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7375..7397
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7435..7453)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7521..7625
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7626..7694,7695..8486)
                     /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(7844..7863)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8657..9245
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9146..9165
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9399..9416
                     /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 catgggcagc agccatcatc atcatcatca cagcagcggc ctggtgccgc
     2521 gcggcagcca tatggcccaa gttgcaatgt ccaccctccc cgttgaagat gaggagtcct
     2581 cggagagcag gatggtggtg acattcctca tgtcagctct cgagtccatg tgtaaagaac
     2641 tggccaagtc caaagccgaa gtggcctgca ttgcagtgta tgaaacagac gtgtttgtcg
     2701 tcggaactga aagaggacgt gcttttgtca ataccagaaa ggattttcaa aaagattttg
     2761 taaaatattg tgttgaagaa gaagaaaaag ctgcagagat gcataaaatg aaatctacaa
     2821 cccaggcaaa tcggatgagt gtagatgctg tagaaattga aacactcaga aaaacagttg
     2881 aggactattt ctgcttttgc tatgggaaag ctttaggcaa atccacagtg gtacctgtac
     2941 catatgagaa gatgctgcga gaccagtcgg ctgtggtagt gcaggggctt ccggaaggtg
     3001 ttgcctttaa acaccccgag aactatgatc ttgcaaccct gaaatggatt ttggagaaca
     3061 aagcaggaat ttcattcatc attaagagac cttttttaga gccaaagaag catgtaggtg
     3121 gtcgtgtgat ggtaacagat gctgacaggt caatactatc tccaggtgga agttgtggcc
     3181 ccatcaaagt gaaaactgaa cccacagaag attctggcat ttccctggaa atggcagctg
     3241 tgacagtaaa ggaagaatca gaagatcctg attattatca atataacatt caaggaagcc
     3301 accattcttc agagggcaat gaaggcacag aaatggaagt accagcagaa gatgatgatt
     3361 attctccacc gtctaagaga ccaaaggcca atgagctacc gcagccacca gtcccggaac
     3421 ccgccaatgc tgggaagcgg aaagtgaggg agttcaactt cgagaaatgg aatgctcgca
     3481 tcactgatct acgtaaacaa gttgaagaat tgtttgaaag gaaatatgct caagccataa
     3541 aagccaaagg tccggtgacg atcccgtacc ctcttttcca gtctcatgtt gaagatcttt
     3601 atgtagaagg acttcctgaa ggaattcctt ttaaaaggcc atctacttac ggaattcctc
     3661 gcctggagag gatattactt gcaaaggaaa ggattcgttt tgtgattaag aaacatgagc
     3721 ttctgaattc aacacgtgaa gatttacagc ttgataagcc agcttcagga gtaaaggaag
     3781 aatggtatgc cagaatcact aaattaagaa agatggtgga tcagcttttc tgcaaaaaat
     3841 ttgcggaacg cttggggagc actgaagcca aggctgtacc gtaccaaaaa tttgaggcac
     3901 acccgaatga tctgtacgtg gaaggactgc cagaaaacat tcctttccga agtccctcat
     3961 ggtatggaat cccaaggctg gaaaaaatca ttcaagtggg caatcgaatt aaatttgtta
     4021 ttaaaagacc agaacttctg actcacagta ccactgaagt tactcagcca agaacgaata
     4081 caccagtcaa agaagattgg aatgtcagaa ttaccaagct acggaagcaa gtggaagaga
     4141 tttttaattt gaaatttgct caagctcttg gactcaccga ggcagtaaaa gtaccatatc
     4201 ctgtgtttga atcaaacccg gagttcttgt atgtggaagg cttgccagag gggattccct
     4261 tccgaagccc tacctggttt ggaattccac gacttgaaag gatcgtccgc gggagtaata
     4321 aaatcaagtt cgttgttaaa aaacctgaac tagttatttc ctacttgcct cctgggatgg
     4381 ctagtaaaat aaacactaaa gctttgcagt cccccaaaag accacgaagt cctgggagta
     4441 attcaaaggt tcctgaaatt gaggtcaccg tggaaggccc taataacaac aatcctcaaa
     4501 cctcagctgt tcgaaccccg acccagacta acggttctaa cgttcccttc aagccacgag
     4561 ggagagagtt ttcctttgag gcctggaatg ccaaaatcac ggacctaaaa cagaaagttg
     4621 aaaatctctt caatgagaaa tgtggggaag ctcttggcct taaacaagct gtgaaggtgc
     4681 cgttcgcgtt atttgagtct ttcccggaag acttttatgt ggaaggctta cctgagggtg
     4741 tgccattccg aagaccatcg acttttggca ttccgagact ggagaagata ctcagaaaca
     4801 aagccaaaat taagttcatc attaaaaagc ccgaaatgtt tgagacggcg attaaggaga
     4861 gcacctcctc taagagccct cccagaaaaa taaattcatc acccaatgtt aatactactg
     4921 catcaggtgt tgaagacctt aacatcattc aggtgacaat tccagatgat gataatgaaa
     4981 gactctcgaa agttgaaaaa gctagacagc taagagaaca agtgaatgac ctctttagtc
     5041 ggaaatttgg tgaagctatt ggtatgggtt ttcctgtgaa agttccctac aggaaaatca
     5101 caattaaccc tggctgtgtg gtggttgatg gcatgccccc gggggtgtcc ttcaaagccc
     5161 ccagctacct ggaaatcagc tccatgagaa ggatcttaga ctctgccgag tttatcaaat
     5221 tcacggtcat tagaccattt ccaggacttg tgattaataa ccagctggtt gatcagagtg
     5281 agtcagaagg ccccgtgata caagaatcag ctgaaccaag ccagttggaa gttccagcca
     5341 cagaagaaat aaaagagact gatggaagct ctcagatcaa gcaagaacca gaccccacgt
     5401 ggtagacctc ttccctccta ggcttaaagt atcagtggtt gagaagagct ttcggacctg
     5461 ttactacccc aagctgtgtg taatatactt gtataacaga aataccttct atacacacct
     5521 ttttttctac ttttagatag aaatgtctac tttttcagca gttctgtgaa ttaaagagca
     5581 gagtgactgt gggtctggaa tggctggtgt acttgggaat gtactatcag gattttacag
     5641 caatgctggg aaatgacagg gaaaatgaca ggaatgaatc tcaccagatt ttttatgtac
     5701 tcagcagagc cttgagttac ggtgtttatt ttccaatcaa gtgaagatat ctcctacttc
     5761 tcctactgga acatctcagc ttctgcagtg aagaaaaatt cctgtgatag ttcagttctt
     5821 tagtttttct atttgaaaaa aaaaaaaaag aattcgatat caagcttatc gatgcggccg
     5881 cgactctaga gtgagggtcc ccacctggga cccttgagag tatcaggtct cccacgtggg
     5941 agacaagaaa tccctgttta atatttaaac agcagtgttc cccatctggg tccttgcacc
     6001 cctcactctg gcctcagccg actgcacagc ggcccctgca tccccttggc tgtgaggccc
     6061 ctggacaagc agaggtggcc agagctggga ggcatggccc tggggtccca cgaatttgct
     6121 ggggaatctc gtttttcttc ttaagacttt tgggacatgg tttgactccc gaacatcacc
     6181 gacgcgtctc ctgtttttct gggtggcctc gggacacctg ccctgccccc acgagggtca
     6241 ggactgtgac tctttttagg gccaggcagg tgcctggaca tttgccttgc tggacgggga
     6301 ctggggatgt gggagggagc agacaggagg aatcatgtca ggcctgtgtg tgaaaggaag
     6361 ctccactgtc accctccacc tcttcacccc ccactcacca gtgtcccctc cactgtcaca
     6421 ttgtaactga acttcaggat aataaagtgt ttgcctccaa aaaaaaaaaa aaaaaaaaaa
     6481 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6541 aaaaaaaaga attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     6601 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     6661 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     6721 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     6781 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6841 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6901 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6961 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     7021 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     7081 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt
     7141 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     7201 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     7261 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     7321 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     7381 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     7441 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     7501 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     7561 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     7621 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     7681 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     7741 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     7801 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     7861 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7921 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7981 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     8041 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     8101 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     8161 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     8221 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     8281 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     8341 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     8401 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     8461 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     8521 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     8581 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     8641 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     8701 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     8761 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     8821 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     8881 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     8941 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     9001 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     9061 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     9121 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     9181 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     9241 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     9301 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     9361 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     9421 c
//
