LOCUS       sequence-10164-e        8296 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 8296)
  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 8296)
  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-p70
FEATURES             Location/Qualifiers
     source          1..8296
                     /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(5430..5447)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5430..5446)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5439..5461)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5659..6114
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5746..5765)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5956..5977
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6131..6150)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6250..6272
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6310..6328)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6396..6500
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6501..6569,6570..7361)
                     /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(6719..6738)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7532..8120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8021..8040
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8274..8291
                     /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 cctgagggtt
     4741 agatcgatgc ggccgcgact ctagagtgag ggtccccacc tgggaccctt gagagtatca
     4801 ggtctcccac gtgggagaca agaaatccct gtttaatatt taaacagcag tgttccccat
     4861 ctgggtcctt gcacccctca ctctggcctc agccgactgc acagcggccc ctgcatcccc
     4921 ttggctgtga ggcccctgga caagcagagg tggccagagc tgggaggcat ggccctgggg
     4981 tcccacgaat ttgctgggga atctcgtttt tcttcttaag acttttggga catggtttga
     5041 ctcccgaaca tcaccgacgc gtctcctgtt tttctgggtg gcctcgggac acctgccctg
     5101 cccccacgag ggtcaggact gtgactcttt ttagggccag gcaggtgcct ggacatttgc
     5161 cttgctggac ggggactggg gatgtgggag ggagcagaca ggaggaatca tgtcaggcct
     5221 gtgtgtgaaa ggaagctcca ctgtcaccct ccacctcttc accccccact caccagtgtc
     5281 ccctccactg tcacattgta actgaacttc aggataataa agtgtttgcc tccaaaaaaa
     5341 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     5401 aaaaaaaaaa aaaaaaaaaa aaagaattca ctggccgtcg ttttacaacg tcgtgactgg
     5461 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     5521 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     5581 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     5641 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5701 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     5761 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5821 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     5881 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5941 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     6001 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     6061 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     6121 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     6181 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     6241 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     6301 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     6361 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6421 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6481 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6541 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6601 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6661 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6721 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6781 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6841 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6901 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6961 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     7021 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     7081 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     7141 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     7201 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     7261 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7321 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     7381 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7441 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7501 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7561 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7621 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     7681 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7741 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7801 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7861 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7921 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7981 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     8041 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     8101 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     8161 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     8221 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     8281 cagtgagcga ggaagc
//
