LOCUS       sequence-10159-e        8945 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 8945)
  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 8945)
  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-deltaNLS1-EGFP
FEATURES             Location/Qualifiers
     source          1..8945
                     /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      4671..4680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4677..4679,4680..4682,4683..5396)
                     /codon_start=1
                     /product="enhanced blue variant of GFP"
                     /label=EBFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4722..4743)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4983..5002)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5330..5351
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6079..6096)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6079..6095)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6088..6110)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6308..6763
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6395..6414)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6605..6626
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6780..6799)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6899..6921
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6959..6977)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7045..7149
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7150..7218,7219..8010)
                     /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(7368..7387)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8181..8769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8670..8689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8923..8940
                     /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 ctaccgcagc
     2641 caccagtccc ggaacccgcc aatgctggga agcggaaagt gagggagttc aacttcgaga
     2701 aatggaatgc tcgcatcact gatctacgta aacaagttga agaattgttt gaaaggaaat
     2761 atgctcaagc cataaaagcc aaaggtccgg tgacgatccc gtaccctctt ttccagtctc
     2821 atgttgaaga tctttatgta gaaggacttc ctgaaggaat tccttttaaa aggccatcta
     2881 cttacggaat tcctcgcctg gagaggatat tacttgcaaa ggaaaggatt cgttttgtga
     2941 ttaagaaaca tgagcttctg aattcaacac gtgaagattt acagcttgat aagccagctt
     3001 caggagtaaa ggaagaatgg tatgccagaa tcactaaatt aagaaagatg gtggatcagc
     3061 ttttctgcaa aaaatttgcg gaacgcttgg ggagcactga agccaaggct gtaccgtacc
     3121 aaaaatttga ggcacacccg aatgatctgt acgtggaagg actgccagaa aacattcctt
     3181 tccgaagtcc ctcatggtat ggaatcccaa ggctggaaaa aatcattcaa gtgggcaatc
     3241 gaattaaatt tgttattaaa agaccagaac ttctgactca cagtaccact gaagttactc
     3301 agccaagaac gaatacacca gtcaaagaag attggaatgt cagaattacc aagctacgga
     3361 agcaagtgga agagattttt aatttgaaat ttgctcaagc tcttggactc accgaggcag
     3421 taaaagtacc atatcctgtg tttgaatcaa acccggagtt cttgtatgtg gaaggcttgc
     3481 cagaggggat tcccttccga agccctacct ggtttggaat tccacgactt gaaaggatcg
     3541 tccgcgggag taataaaatc aagttcgttg ttaaaaaacc tgaactagtt atttcctact
     3601 tgcctcctgg gatggctagt aaaataaaca ctaaagcttt gcagtccccc aaaagaccac
     3661 gaagtcctgg gagtaattca aaggttcctg aaattgaggt caccgtggaa ggccctaata
     3721 acaacaatcc tcaaacctca gctgttcgaa ccccgaccca gactaacggt tctaacgttc
     3781 ccttcaagcc acgagggaga gagttttcct ttgaggcctg gaatgccaaa atcacggacc
     3841 taaaacagaa agttgaaaat ctcttcaatg agaaatgtgg ggaagctctt ggccttaaac
     3901 aagctgtgaa ggtgccgttc gcgttatttg agtctttccc ggaagacttt tatgtggaag
     3961 gcttacctga gggtgtgcca ttccgaagac catcgacttt tggcattccg agactggaga
     4021 agatactcag aaacaaagcc aaaattaagt tcatcattaa aaagcccgaa atgtttgaga
     4081 cggcgattaa ggagagcacc tcctctaaga gccctcccag aaaaataaat tcatcaccca
     4141 atgttaatac tactgcatca ggtgttgaag accttaacat cattcaggtg acaattccag
     4201 atgatgataa tgaaagactc tcgaaagttg aaaaagctag acagctaaga gaacaagtga
     4261 atgacctctt tagtcggaaa tttggtgaag ctattggtat gggttttcct gtgaaagttc
     4321 cctacaggaa aatcacaatt aaccctggct gtgtggtggt tgatggcatg cccccggggg
     4381 tgtccttcaa agcccccagc tacctggaaa tcagctccat gagaaggatc ttagactctg
     4441 ccgagtttat caaattcacg gtcattagac catttccagg acttgtgatt aataaccagc
     4501 tggttgatca gagtgagtca gaaggccccg tgatacaaga atcagctgaa ccaagccagt
     4561 tggaagttcc agccacagaa gaaataaaag agactgatgg aagctctcag atcaagcaag
     4621 aaccagaccc cacgtggtcg acggtaccgc gggcccggga tccaccggtc gccaccatgg
     4681 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     4741 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     4801 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     4861 tgaccaccct gacccacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     4921 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     4981 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     5041 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     5101 tggagtacaa cttcaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     5161 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     5221 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     5281 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     5341 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     5401 gccgcgactc tagagtgagg gtccccacct gggacccttg agagtatcag gtctcccacg
     5461 tgggagacaa gaaatccctg tttaatattt aaacagcagt gttccccatc tgggtccttg
     5521 cacccctcac tctggcctca gccgactgca cagcggcccc tgcatcccct tggctgtgag
     5581 gcccctggac aagcagaggt ggccagagct gggaggcatg gccctggggt cccacgaatt
     5641 tgctggggaa tctcgttttt cttcttaaga cttttgggac atggtttgac tcccgaacat
     5701 caccgacgcg tctcctgttt ttctgggtgg cctcgggaca cctgccctgc ccccacgagg
     5761 gtcaggactg tgactctttt tagggccagg caggtgcctg gacatttgcc ttgctggacg
     5821 gggactgggg atgtgggagg gagcagacag gaggaatcat gtcaggcctg tgtgtgaaag
     5881 gaagctccac tgtcaccctc cacctcttca ccccccactc accagtgtcc cctccactgt
     5941 cacattgtaa ctgaacttca ggataataaa gtgtttgcct ccaaaaaaaa aaaaaaaaaa
     6001 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6061 aaaaaaaaaa aagaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     6121 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     6181 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     6241 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     6301 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     6361 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     6421 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6481 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     6541 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6601 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt
     6661 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6721 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     6781 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     6841 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     6901 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     6961 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     7021 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     7081 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     7141 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     7201 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     7261 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     7321 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     7381 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     7441 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     7501 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     7561 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     7621 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     7681 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     7741 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     7801 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7861 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7921 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7981 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     8041 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     8101 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     8161 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     8221 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     8281 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     8341 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     8401 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     8461 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     8521 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     8581 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     8641 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     8701 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     8761 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8821 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     8881 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     8941 gaagc
//
