LOCUS       sequence-10160-e        9397 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 9397)
  AUTHORS   Cheriyath V, Roy AL
  TITLE     Alternatively spliced isoforms of TFII-I. Complex formation, nuclear
            translocation, and differential gene regulation.
  JOURNAL   J Biol Chem. 2000 Aug 25. 275(34):26300-8.
  PUBMED    10854432
REFERENCE   2  (bases 1 to 9397)
  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. 2000 Aug 25. 275(34):26300-8."
COMMENT     Sequence Label: pEBG-TFII-deltaNLS1
FEATURES             Location/Qualifiers
     source          1..9397
                     /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(6531..6548)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6531..6547)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6540..6562)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6760..7215
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6847..6866)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7057..7078
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7232..7251)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7351..7373
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7411..7429)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7497..7601
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7602..7670,7671..8462)
                     /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(7820..7839)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8633..9221
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9122..9141
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9375..9392
                     /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 attctaatga gctaccgcag ccaccagtcc cggaacccgc caatgctggg aagcggaaag
     3421 tgagggagtt caacttcgag aaatggaatg ctcgcatcac tgatctacgt aaacaagttg
     3481 aagaattgtt tgaaaggaaa tatgctcaag ccataaaagc caaaggtccg gtgacgatcc
     3541 cgtaccctct tttccagtct catgttgaag atctttatgt agaaggactt cctgaaggaa
     3601 ttccttttaa aaggccatct acttacggaa ttcctcgcct ggagaggata ttacttgcaa
     3661 aggaaaggat tcgttttgtg attaagaaac atgagcttct gaattcaaca cgtgaagatt
     3721 tacagcttga taagccagct tcaggagtaa aggaagaatg gtatgccaga atcactaaat
     3781 taagaaagat ggtggatcag cttttctgca aaaaatttgc ggaacgcttg gggagcactg
     3841 aagccaaggc tgtaccgtac caaaaatttg aggcacaccc gaatgatctg tacgtggaag
     3901 gactgccaga aaacattcct ttccgaagtc cctcatggta tggaatccca aggctggaaa
     3961 aaatcattca agtgggcaat cgaattaaat ttgttattaa aagaccagaa cttctgactc
     4021 acagtaccac tgaagttact cagccaagaa cgaatacacc agtcaaagaa gattggaatg
     4081 tcagaattac caagctacgg aagcaagtgg aagagatttt taatttgaaa tttgctcaag
     4141 ctcttggact caccgaggca gtaaaagtac catatcctgt gtttgaatca aacccggagt
     4201 tcttgtatgt ggaaggcttg ccagagggga ttcccttccg aagccctacc tggtttggaa
     4261 ttccacgact tgaaaggatc gtccgcggga gtaataaaat caagttcgtt gttaaaaaac
     4321 ctgaactagt tatttcctac ttgcctcctg ggatggctag taaaataaac actaaagctt
     4381 tgcagtcccc caaaagacca cgaagtcctg ggagtaattc aaaggttcct gaaattgagg
     4441 tcaccgtgga aggccctaat aacaacaatc ctcaaacctc agctgttcga accccgaccc
     4501 agactaacgg ttctaacgtt cccttcaagc cacgagggag agagttttcc tttgaggcct
     4561 ggaatgccaa aatcacggac ctaaaacaga aagttgaaaa tctcttcaat gagaaatgtg
     4621 gggaagctct tggccttaaa caagctgtga aggtgccgtt cgcgttattt gagtctttcc
     4681 cggaagactt ttatgtggaa ggcttacctg agggtgtgcc attccgaaga ccatcgactt
     4741 ttggcattcc gagactggag aagatactca gaaacaaagc caaaattaag ttcatcatta
     4801 aaaagcccga aatgtttgag acggcgatta aggagagcac ctcctctaag agccctccca
     4861 gaaaaataaa ttcatcaccc aatgttaata ctactgcatc aggtgttgaa gaccttaaca
     4921 tcattcaggt gacaattcca gatgatgata atgaaagact ctcgaaagtt gaaaaagcta
     4981 gacagctaag agaacaagtg aatgacctct ttagtcggaa atttggtgaa gctattggta
     5041 tgggttttcc tgtgaaagtt ccctacagga aaatcacaat taaccctggc tgtgtggtgg
     5101 ttgatggcat gcccccgggg gtgtccttca aagcccccag ctacctggaa atcagctcca
     5161 tgagaaggat cttagactct gccgagttta tcaaattcac ggtcattaga ccatttccag
     5221 gacttgtgat taataaccag ctggttgatc agagtgagtc agaaggcccc gtgatacaag
     5281 aatcagctga accaagccag ttggaagttc cagccacaga agaaataaaa gagactgatg
     5341 gaagctctca gatcaagcaa gaaccagacc ccacgtggta gacctcttcc ctcctaggct
     5401 taaagtatca gtggttgaga agagctttcg gacctgttac taccccaagc tgtgtgtaat
     5461 atacttgtat aacagaaata ccttctatac acaccttttt ttctactttt agatagaaat
     5521 gtctactttt tcagcagttc tgtgaattaa agagcagagt gactgtgggt ctggaatggc
     5581 tggtgtactt gggaatgtac tatcaggatt ttacagcaat gctgggaaat gacagggaaa
     5641 atgacaggaa tgaatctcac cagatttttt atgtactcag cagagccttg agttacggtg
     5701 tttattttcc aatcaagtga agatatctcc tacttctcct actggaacat ctcagcttct
     5761 gcagtgaaga aaaattcctg tgatagttca gttctttagt ttttctattt gaaaaaaaaa
     5821 aaaaagaatt cgatatcaag cttatcgatg cggccgcgac tctagagtga gggtccccac
     5881 ctgggaccct tgagagtatc aggtctccca cgtgggagac aagaaatccc tgtttaatat
     5941 ttaaacagca gtgttcccca tctgggtcct tgcacccctc actctggcct cagccgactg
     6001 cacagcggcc cctgcatccc cttggctgtg aggcccctgg acaagcagag gtggccagag
     6061 ctgggaggca tggccctggg gtcccacgaa tttgctgggg aatctcgttt ttcttcttaa
     6121 gacttttggg acatggtttg actcccgaac atcaccgacg cgtctcctgt ttttctgggt
     6181 ggcctcggga cacctgccct gcccccacga gggtcaggac tgtgactctt tttagggcca
     6241 ggcaggtgcc tggacatttg ccttgctgga cggggactgg ggatgtggga gggagcagac
     6301 aggaggaatc atgtcaggcc tgtgtgtgaa aggaagctcc actgtcaccc tccacctctt
     6361 caccccccac tcaccagtgt cccctccact gtcacattgt aactgaactt caggataata
     6421 aagtgtttgc ctccaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6481 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaagaattc actggccgtc
     6541 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     6601 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     6661 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     6721 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     6781 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     6841 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     6901 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     6961 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     7021 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     7081 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     7141 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     7201 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     7261 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     7321 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     7381 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     7441 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     7501 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     7561 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     7621 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     7681 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     7741 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     7801 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     7861 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     7921 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     7981 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     8041 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     8101 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     8161 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     8221 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     8281 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     8341 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     8401 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     8461 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     8521 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     8581 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     8641 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     8701 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     8761 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     8821 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     8881 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     8941 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     9001 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     9061 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     9121 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     9181 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     9241 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     9301 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     9361 gaacgaccga gcgcagcgag tcagtgagcg aggaagc
//
