LOCUS       sequence-10111-e        8911 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 8911)
  AUTHORS   Tussie-Luna MI, Bayarsaihan D, Ruddle FH, Roy AL
  TITLE     Repression of TFII-I-dependent transcription by nuclear exclusion.
  JOURNAL   Proc Natl Acad Sci U S A. 2001 Jul 3. 98(14):7789-94.
  PUBMED    11438732
REFERENCE   2  (bases 1 to 8911)
  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: "Proc Natl 
            Acad Sci U S A. 2001 Jul 3. 98(14):7789-94."
COMMENT     Sequence Label: pEBB-GFP-hBEN
FEATURES             Location/Qualifiers
     source          1..8911
                     /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             join(1805..1807,1808..1810,1811..2521)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1805..1807,1808..1810,1811..2521)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1850..1871)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2111..2130)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2458..2479
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6045..6062)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6045..6061)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6054..6076)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6274..6729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6361..6380)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6571..6592
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6746..6765)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6865..6887
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6925..6943)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7011..7115
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7116..7184,7185..7976)
                     /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(7334..7353)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8147..8735
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8636..8655
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8889..8906
                     /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 ggatccacta
     1801 gtccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1861 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1921 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1981 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2041 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2101 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2161 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2221 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2281 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2341 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2401 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     2461 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2521 gatcgatgga atggccttgc tgggtaagcg ctgtgacgtc cccaccaacg gctgcggacc
     2581 cgaccgctgg aactccgcgt tcacccgcaa agacgagatc atcaccagcc tcgtgtctgc
     2641 cttagactcc atgtgctcag cgctgtccaa actgaacgcc gaggtggcct gtgtcgccgt
     2701 gcacgatgag agcgcctttg tggtgggcac agagaagggg agaatgttcc tgaatgcccg
     2761 gaaggagcta cagtcagact tcctcaggtt ctgccgaggg cccccgtgga aggatccgga
     2821 ggcagagcac cccaagaagg tgcagcgggg cgagggtgga agccgtagcc tccctcggtc
     2881 ctccctggaa catggctcag atgtgtacct tctgcggaag atggtagagg aggtgtttga
     2941 tgttctttat agcgaggccc tgggaagggc cagtgtggtg ccactgccct atgagaggct
     3001 gctcagggag ccagggctgc tggccgtgca ggggctgccc gaaggcctgg ccttccgaag
     3061 gccagccgag tatgacccca aggccctcat ggccatcctg gaacacagcc accgcatccg
     3121 cttcaagctc aagaggccac ttgaggatgg cgggcgggac tcgaaggccc tggtggagct
     3181 gaacggtgtc tccctgattc ccaaggggtc acgggactgt ggcctgcatg gccaggcccc
     3241 caaggtgcca ccccaggacc tgcccccaac cgccacctcc tcctccatgg ccagcttcct
     3301 gtacagcacg gcgctcccca accacgccat ccgagagctc aagcaggaag caccttcctg
     3361 cccccttgcc cccagcgacc tgggcctgag tcggcccatg ccagagccca aggccaccgg
     3421 tgcccaagac ttctccgact gttgtggaca gaagcccact gggcctggtg ggcctctcat
     3481 ccagaacgtc catgcctcca agcgcattct cttctccatc gtccatgaca agtcagagaa
     3541 gtgggacgcc ttcataaagg aaaccgagga catcaacacg ctccgggagt gtgtgcagat
     3601 cctgtttaac agcagatatg cggaagccct gggcctggac cacatggtcc ccgtgcccta
     3661 ccggaagatt gcctgtgacc cggaggctgt ggagatcgtg ggcatcccgg acaagatccc
     3721 cttcaagcgc ccctgcactt atggagtccc caagctgaag cggatcctgg aggagcgcca
     3781 tagtatccac ttcatcatta agaggatgtt tgatgagcga attttcacag ggaacaagtt
     3841 taccaaagac accacgaagc tggagccagc cagcccgcca gaggacacct ctgcagaggt
     3901 ctctagggcc accgtccttg accttgctgg gaatgctcgg tcagacaagg gcagcatgtc
     3961 tgaagactgt gggccaggaa cgtccgggga gctgggcggg ctgaggccga tcaaaattga
     4021 gccagaggat ctggacatca ttcaggtcac cgtcccagac ccctcgccaa cctctgagga
     4081 aatgacagac tcgatgcctg ggcacctgcc atcggaggat tctggttatg ggatggagat
     4141 gctgacagac aaaggtctga gtgaggacgc gcggccggag gagaggcccg tggaggacag
     4201 ccacggtgac gtgatccggc ccctgcggaa gcaggtggag ctgctcttca acacacgata
     4261 cgccaaggcc attggcatct cggagcccgt caaggtgccg tactccaagt ttctgatgca
     4321 cccggaggag ctgtttgtgg tgggactgcc tgaaggcatc tccctccgca ggcccaactg
     4381 cttcgggatc gccaagctcc ggaagattct ggaggccagc aacagcatcc agtttgtcat
     4441 caagaggccc gagctgctca ctgagggagt caaagagccc atcatggata gtcaagagag
     4501 ggattccggg gaccctctgg tggacgagag cctgaagaga cagggctttc aagaaaatta
     4561 tgacgcgagg ctctcacgga tcgacatcgc caacacacta agggagcagg tccaggacct
     4621 tttcaataag aaatacgggg aagccttggg catcaagtac ccggtccagg tcccctacaa
     4681 gcggatcaag agtaaccccg gctccgtgat catcgagggg ctgcccccag gaatcccgtt
     4741 ccgaaagccc tgtaccttcg gctcccagaa cctggagagg attcttgctg tggctgacaa
     4801 gatcaagttc acagtcacca ggcctttcca aggactcatc ccaaagcctg atgaagatga
     4861 cgccaacaga ctcggggaga aggtgatcct gcgggagcag gtgaaggaac tcttcaacga
     4921 gaaatacggt gaggccctgg gcctgaaccg gccggtgctg gtcccttata aactaatccg
     4981 ggacagccca gacgccgtgg aggtcacggg tctgcctgat gacatcccct tccggaaccc
     5041 caacacgtac gacatccacc ggctggagaa gatcctgaag gcccgagagc atgtccgcat
     5101 ggtcatcatt aaccagctcc aaccctttgc agaaatctgc aatgatgcca aggtgccagc
     5161 caaagacagc agcattccca agcgcaagag aaagcgggtc tcggaaggaa attccgtctc
     5221 ctcttcctcc tcgtcttcct cttcctcgtc ctctaacccg gattcagtgg catcggccaa
     5281 ccagatctca ctcgtgcaat ggccaatgta catggtggac tatgccggcc tgaacgtgca
     5341 gctcccggga cctcttaatt acagcggccg cgactctaga gtgagggtcc ccacctggga
     5401 cccttgagag tatcaggtct cccacgtggg agacaagaaa tccctgttta atatttaaac
     5461 agcagtgttc cccatctggg tccttgcacc cctcactctg gcctcagccg actgcacagc
     5521 ggcccctgca tccccttggc tgtgaggccc ctggacaagc agaggtggcc agagctggga
     5581 ggcatggccc tggggtccca cgaatttgct ggggaatctc gtttttcttc ttaagacttt
     5641 tgggacatgg tttgactccc gaacatcacc gacgcgtctc ctgtttttct gggtggcctc
     5701 gggacacctg ccctgccccc acgagggtca ggactgtgac tctttttagg gccaggcagg
     5761 tgcctggaca tttgccttgc tggacgggga ctggggatgt gggagggagc agacaggagg
     5821 aatcatgtca ggcctgtgtg tgaaaggaag ctccactgtc accctccacc tcttcacccc
     5881 ccactcacca gtgtcccctc cactgtcaca ttgtaactga acttcaggat aataaagtgt
     5941 ttgcctccaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6001 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaga attcactggc cgtcgtttta
     6061 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     6121 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     6181 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     6241 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     6301 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     6361 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6421 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     6481 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6541 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     6601 tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     6661 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     6721 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     6781 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     6841 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     6901 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     6961 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     7021 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     7081 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     7141 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     7201 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     7261 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     7321 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     7381 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     7441 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     7501 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     7561 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     7621 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     7681 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     7741 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     7801 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     7861 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7921 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7981 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     8041 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     8101 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     8161 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     8221 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     8281 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     8341 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     8401 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8461 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     8521 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     8581 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     8641 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     8701 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     8761 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     8821 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     8881 ccgagcgcag cgagtcagtg agcgaggaag c
//
