LOCUS       sequence-9954-ex        8975 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 8975)
  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 8975)
  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: pEBB-II-I-EGFP delta isoform
FEATURES             Location/Qualifiers
     source          1..8975
                     /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"
     regulatory      4701..4710
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4707..4709,4710..4712,4713..5426)
                     /codon_start=1
                     /product="enhanced blue variant of GFP"
                     /label=EBFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4752..4773)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5013..5032)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5360..5381
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6109..6126)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6109..6125)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6118..6140)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6338..6793
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6425..6444)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6635..6656
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6810..6829)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6929..6951
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6989..7007)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7075..7179
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7180..7248,7249..8040)
                     /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(7398..7417)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8211..8799
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8700..8719
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8953..8970
                     /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 ggatccatgg
     1801 cccaagttgc aatgtccacc ctccccgttg aagatgagga gtcctcggag agcaggatgg
     1861 tggtgacatt cctcatgtca gctctcgagt ccatgtgtaa agaactggcc aagtccaaag
     1921 ccgaagtggc ctgcattgca gtgtatgaaa cagacgtgtt tgtcgtcgga actgaaagag
     1981 gacgtgcttt tgtcaatacc agaaaggatt ttcaaaaaga ttttgtaaaa tattgtgttg
     2041 aagaagaaga aaaagctgca gagatgcata aaatgaaatc tacaacccag gcaaatcgga
     2101 tgagtgtaga tgctgtagaa attgaaacac tcagaaaaac agttgaggac tatttctgct
     2161 tttgctatgg gaaagcttta ggcaaatcca cagtggtacc tgtaccatat gagaagatgc
     2221 tgcgagacca gtcggctgtg gtagtgcagg ggcttccgga aggtgttgcc tttaaacacc
     2281 ccgagaacta tgatcttgca accctgaaat ggattttgga gaacaaagca ggaatttcat
     2341 tcatcattaa gagacctttt ttagagccaa agaagcatgt aggtggtcgt gtgatggtaa
     2401 cagatgctga caggtcaata ctatctccag gtggaagttg tggccccatc aaagtgaaaa
     2461 ctgaacccac agaagattct ggcatttccc tggaaatggc agctgtgaca gtaaaggaag
     2521 aatcagaaga tcctgattat tatcaatata acattcaagg aagccaccat tcttcagagg
     2581 gcaatgaagg cacagaaatg gaagtaccag cagaagatga tgattattct ccaccgtcta
     2641 agagaccaaa ggccaatgag ctaccgcagc caccagtccc ggaacccgcc aatgctggga
     2701 agcggaaagt gagggagttc aacttcgaga aatggaatgc tcgcatcact gatctacgta
     2761 aacaagttga agaattgttt gaaaggaaat atgctcaagc cataaaagcc aaaggtccgg
     2821 tgacgatccc gtaccctctt ttccagtctc atgttgaaga tctttatgta gaaggacttc
     2881 ctgaaggaat tccttttaaa aggccatcta cttacggaat tcctcgcctg gagaggatat
     2941 tacttgcaaa ggaaaggatt cgttttgtga ttaagaaaca tgagcttctg aattcaacac
     3001 gtgaagattt acagcttgat aagccagctt caggagtaaa ggaagaatgg tatgccagaa
     3061 tcactaaatt aagaaagatg gtggatcagc ttttctgcaa aaaatttgcg gaacgcttgg
     3121 ggagcactga agccaaggct gtaccgtacc aaaaatttga ggcacacccg aatgatctgt
     3181 acgtggaagg actgccagaa aacattcctt tccgaagtcc ctcatggtat ggaatcccaa
     3241 ggctggaaaa aatcattcaa gtgggcaatc gaattaaatt tgttattaaa agaccagaac
     3301 ttctgactca cagtaccact gaagttactc agccaagaac gaatacacca gtcaaagaag
     3361 attggaatgt cagaattacc aagctacgga agcaagtgga agagattttt aatttgaaat
     3421 ttgctcaagc tcttggactc accgaggcag taaaagtacc atatcctgtg tttgaatcaa
     3481 acccggagtt cttgtatgtg gaaggcttgc cagaggggat tcccttccga agccctacct
     3541 ggtttggaat tccacgactt gaaaggatcg tccgcgggag taataaaatc aagttcgttg
     3601 ttaaaaaacc tgaactagtt atttcctact tgcctcctgg gatggctagt aaaataaaca
     3661 ctaaagcttt gcagtccccc aaaagaccac gaagtcctgg gagtaattca aaggttcctg
     3721 aaattgaggt caccgtggaa ggccctaata acaacaatcc tcaaacctca gctgttcgaa
     3781 ccccgaccca gactaacggt tctaacgttc ccttcaagcc acgagggaga gagttttcct
     3841 ttgaggcctg gaatgccaaa atcacggacc taaaacagaa agttgaaaat ctcttcaatg
     3901 agaaatgtgg ggaagctctt ggccttaaac aagctgtgaa ggtgccgttc gcgttatttg
     3961 agtctttccc ggaagacttt tatgtggaag gcttacctga gggtgtgcca ttccgaagac
     4021 catcgacttt tggcattccg agactggaga agatactcag aaacaaagcc aaaattaagt
     4081 tcatcattaa aaagcccgaa atgtttgaga cggcgattaa ggagagcacc tcctctaaga
     4141 gccctcccag aaaaataaat tcatcaccca atgttaatac tactgcatca ggtgttgaag
     4201 accttaacat cattcaggtg acaattccag atgatgataa tgaaagactc tcgaaagttg
     4261 aaaaagctag acagctaaga gaacaagtga atgacctctt tagtcggaaa tttggtgaag
     4321 ctattggtat gggttttcct gtgaaagttc cctacaggaa aatcacaatt aaccctggct
     4381 gtgtggtggt tgatggcatg cccccggggg tgtccttcaa agcccccagc tacctggaaa
     4441 tcagctccat gagaaggatc ttagactctg ccgagtttat caaattcacg gtcattagac
     4501 catttccagg acttgtgatt aataaccagc tggttgatca gagtgagtca gaaggccccg
     4561 tgatacaaga atcagctgaa ccaagccagt tggaagttcc agccacagaa gaaataaaag
     4621 agactgatgg aagctctcag atcaagcaag aaccagaccc cacgtggtcg acggtaccgc
     4681 gggcccggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     4741 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     4801 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     4861 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacccacggc gtgcagtgct
     4921 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     4981 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     5041 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     5101 aggaggacgg caacatcctg gggcacaagc tggagtacaa cttcaacagc cacaacgtct
     5161 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     5221 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     5281 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     5341 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     5401 tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagagtgagg gtccccacct
     5461 gggacccttg agagtatcag gtctcccacg tgggagacaa gaaatccctg tttaatattt
     5521 aaacagcagt gttccccatc tgggtccttg cacccctcac tctggcctca gccgactgca
     5581 cagcggcccc tgcatcccct tggctgtgag gcccctggac aagcagaggt ggccagagct
     5641 gggaggcatg gccctggggt cccacgaatt tgctggggaa tctcgttttt cttcttaaga
     5701 cttttgggac atggtttgac tcccgaacat caccgacgcg tctcctgttt ttctgggtgg
     5761 cctcgggaca cctgccctgc ccccacgagg gtcaggactg tgactctttt tagggccagg
     5821 caggtgcctg gacatttgcc ttgctggacg gggactgggg atgtgggagg gagcagacag
     5881 gaggaatcat gtcaggcctg tgtgtgaaag gaagctccac tgtcaccctc cacctcttca
     5941 ccccccactc accagtgtcc cctccactgt cacattgtaa ctgaacttca ggataataaa
     6001 gtgtttgcct ccaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6061 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aagaattcac tggccgtcgt
     6121 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     6181 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     6241 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     6301 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     6361 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     6421 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     6481 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     6541 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     6601 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     6661 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     6721 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     6781 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     6841 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     6901 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     6961 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     7021 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     7081 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     7141 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     7201 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     7261 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     7321 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     7381 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     7441 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7501 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     7561 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     7621 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7681 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     7741 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7801 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7861 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7921 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7981 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     8041 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     8101 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     8161 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     8221 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     8281 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     8341 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     8401 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     8461 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     8521 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     8581 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     8641 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     8701 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     8761 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     8821 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     8881 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     8941 acgaccgagc gcagcgagtc agtgagcgag gaagc
//
