LOCUS       sequence-10163-e        9151 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 9151)
  AUTHORS   Cheriyath V, Roy AL
  TITLE     Structure-function analysis of TFII-I. Roles of the N-terminal end, 
            basic region, and I-repeats.
  JOURNAL   J Biol Chem. 2001 Mar 16. 276(11):8377-83.
  PUBMED    11113127
REFERENCE   2  (bases 1 to 9151)
  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: "J Biol 
            Chem. 2001 Mar 16. 276(11):8377-83."
COMMENT     Sequence Label: pEBG-TFII-I N90
FEATURES             Location/Qualifiers
     source          1..9151
                     /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(6285..6302)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6285..6301)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6294..6316)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6514..6969
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6601..6620)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6811..6832
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6986..7005)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7105..7127
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7165..7183)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7251..7355
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7356..7424,7425..8216)
                     /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(7574..7593)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8387..8975
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8876..8895
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9129..9146
                     /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 tcataaaatg aaatctacaa cccaggcaaa tcggatgagt gtagatgctg
     2581 tagaaattga aacactcaga aaaacagttg aggactattt ctgcttttgc tatgggaaag
     2641 ctttaggcaa atccacagtg gtacctgtac catatgagaa gatgctgcga gaccagtcgg
     2701 ctgtggtagt gcaggggctt ccggaaggtg ttgcctttaa acaccccgag aactatgatc
     2761 ttgcaaccct gaaatggatt ttggagaaca aagcaggaat ttcattcatc attaagagac
     2821 cttttttaga gccaaagaag catgtaggtg gtcgtgtgat ggtaacagat gctgacaggt
     2881 caatactatc tccaggtgga agttgtggcc ccatcaaagt gaaaactgaa cccacagaag
     2941 attctggcat ttccctggaa atggcagctg tgacagtaaa ggaagaatca gaagatcctg
     3001 attattatca atataacatt caaggaagcc accattcttc agagggcaat gaaggcacag
     3061 aaatggaagt accagcagaa gatgatgatt attctccacc gtctaagaga ccaaaggcca
     3121 atgagctacc gcagccacca gtcccggaac ccgccaatgc tgggaagcgg aaagtgaggg
     3181 agttcaactt cgagaaatgg aatgctcgca tcactgatct acgtaaacaa gttgaagaat
     3241 tgtttgaaag gaaatatgct caagccataa aagccaaagg tccggtgacg atcccgtacc
     3301 ctcttttcca gtctcatgtt gaagatcttt atgtagaagg acttcctgaa ggaattcctt
     3361 ttaaaaggcc atctacttac ggaattcctc gcctggagag gatattactt gcaaaggaaa
     3421 ggattcgttt tgtgattaag aaacatgagc ttctgaattc aacacgtgaa gatttacagc
     3481 ttgataagcc agcttcagga gtaaaggaag aatggtatgc cagaatcact aaattaagaa
     3541 agatggtgga tcagcttttc tgcaaaaaat ttgcggaacg cttggggagc actgaagcca
     3601 aggctgtacc gtaccaaaaa tttgaggcac acccgaatga tctgtacgtg gaaggactgc
     3661 cagaaaacat tcctttccga agtccctcat ggtatggaat cccaaggctg gaaaaaatca
     3721 ttcaagtggg caatcgaatt aaatttgtta ttaaaagacc agaacttctg actcacagta
     3781 ccactgaagt tactcagcca agaacgaata caccagtcaa agaagattgg aatgtcagaa
     3841 ttaccaagct acggaagcaa gtggaagaga tttttaattt gaaatttgct caagctcttg
     3901 gactcaccga ggcagtaaaa gtaccatatc ctgtgtttga atcaaacccg gagttcttgt
     3961 atgtggaagg cttgccagag gggattccct tccgaagccc tacctggttt ggaattccac
     4021 gacttgaaag gatcgtccgc gggagtaata aaatcaagtt cgttgttaaa aaacctgaac
     4081 tagttatttc ctacttgcct cctgggatgg ctagtaaaat aaacactaaa gctttgcagt
     4141 cccccaaaag accacgaagt cctgggagta attcaaaggt tcctgaaatt gaggtcaccg
     4201 tggaaggccc taataacaac aatcctcaaa cctcagctgt tcgaaccccg acccagacta
     4261 acggttctaa cgttcccttc aagccacgag ggagagagtt ttcctttgag gcctggaatg
     4321 ccaaaatcac ggacctaaaa cagaaagttg aaaatctctt caatgagaaa tgtggggaag
     4381 ctcttggcct taaacaagct gtgaaggtgc cgttcgcgtt atttgagtct ttcccggaag
     4441 acttttatgt ggaaggctta cctgagggtg tgccattccg aagaccatcg acttttggca
     4501 ttccgagact ggagaagata ctcagaaaca aagccaaaat taagttcatc attaaaaagc
     4561 ccgaaatgtt tgagacggcg attaaggaga gcacctcctc taagagccct cccagaaaaa
     4621 taaattcatc acccaatgtt aatactactg catcaggtgt tgaagacctt aacatcattc
     4681 aggtgacaat tccagatgat gataatgaaa gactctcgaa agttgaaaaa gctagacagc
     4741 taagagaaca agtgaatgac ctctttagtc ggaaatttgg tgaagctatt ggtatgggtt
     4801 ttcctgtgaa agttccctac aggaaaatca caattaaccc tggctgtgtg gtggttgatg
     4861 gcatgccccc gggggtgtcc ttcaaagccc ccagctacct ggaaatcagc tccatgagaa
     4921 ggatcttaga ctctgccgag tttatcaaat tcacggtcat tagaccattt ccaggacttg
     4981 tgattaataa ccagctggtt gatcagagtg agtcagaagg ccccgtgata caagaatcag
     5041 ctgaaccaag ccagttggaa gttccagcca cagaagaaat aaaagagact gatggaagct
     5101 ctcagatcaa gcaagaacca gaccccacgt ggtagacctc ttccctccta ggcttaaagt
     5161 atcagtggtt gagaagagct ttcggacctg ttactacccc aagctgtgtg taatatactt
     5221 gtataacaga aataccttct atacacacct ttttttctac ttttagatag aaatgtctac
     5281 tttttcagca gttctgtgaa ttaaagagca gagtgactgt gggtctggaa tggctggtgt
     5341 acttgggaat gtactatcag gattttacag caatgctggg aaatgacagg gaaaatgaca
     5401 ggaatgaatc tcaccagatt ttttatgtac tcagcagagc cttgagttac ggtgtttatt
     5461 ttccaatcaa gtgaagatat ctcctacttc tcctactgga acatctcagc ttctgcagtg
     5521 aagaaaaatt cctgtgatag ttcagttctt tagtttttct atttgaaaaa aaaaaaaaag
     5581 aattcgatat caagcttatc gatgcggccg cgactctaga gtgagggtcc ccacctggga
     5641 cccttgagag tatcaggtct cccacgtggg agacaagaaa tccctgttta atatttaaac
     5701 agcagtgttc cccatctggg tccttgcacc cctcactctg gcctcagccg actgcacagc
     5761 ggcccctgca tccccttggc tgtgaggccc ctggacaagc agaggtggcc agagctggga
     5821 ggcatggccc tggggtccca cgaatttgct ggggaatctc gtttttcttc ttaagacttt
     5881 tgggacatgg tttgactccc gaacatcacc gacgcgtctc ctgtttttct gggtggcctc
     5941 gggacacctg ccctgccccc acgagggtca ggactgtgac tctttttagg gccaggcagg
     6001 tgcctggaca tttgccttgc tggacgggga ctggggatgt gggagggagc agacaggagg
     6061 aatcatgtca ggcctgtgtg tgaaaggaag ctccactgtc accctccacc tcttcacccc
     6121 ccactcacca gtgtcccctc cactgtcaca ttgtaactga acttcaggat aataaagtgt
     6181 ttgcctccaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6241 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaga attcactggc cgtcgtttta
     6301 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     6361 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     6421 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     6481 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     6541 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     6601 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6661 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     6721 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6781 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     6841 tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     6901 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     6961 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     7021 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     7081 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     7141 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     7201 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     7261 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     7321 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     7381 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     7441 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     7501 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     7561 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     7621 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     7681 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     7741 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     7801 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     7861 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     7921 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     7981 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     8041 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     8101 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     8161 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     8221 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     8281 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     8341 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     8401 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     8461 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     8521 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     8581 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     8641 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8701 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     8761 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     8821 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     8881 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     8941 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     9001 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     9061 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     9121 ccgagcgcag cgagtcagtg agcgaggaag c
//
