LOCUS       sequence-339920-        9720 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 9720)
  AUTHORS   Xi Q, He W, Zhang XH, Le HV, Massague J
  TITLE     Genome-wide impact of the BRG1 SWI/SNF chromatin remodeler on the 
            transforming growth factor beta transcriptional program.
  JOURNAL   J Biol Chem. 2008 Jan 11. 283(2):1146-55.
  PUBMED    18003620
REFERENCE   2  (bases 1 to 9720)
  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. 2008 Jan 11. 283(2):1146-55."
COMMENT     Sequence Label: pCMV5 BRGI-Flag
FEATURES             Location/Qualifiers
     source          1..9720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     70..92
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     84..101
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     85..101
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        262..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          940..1036
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             5977..6000
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    6026..6648
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6225..6244)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6672..6692)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6837..6971
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7023..7042)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7024..7042)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7056..7072)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7056..7072)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7069..7091)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7080..7096
                     /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)."
     promoter        complement(join(7104..7110,7111..7128,7129..7134))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7149..7170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7287..7304)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7458..8046)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7538..7557)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8217..9008,9009..9077))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIYYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8840..8859
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9078..9182)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9209..9664
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9296..9315)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9506..9527
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
       61 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg atctatacat
      121 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg
      181 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat
      241 gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctcgttt agtgaaccgt cagaattcag atctggtacc acgcgtatcg ataagcttgc
      901 atgcctgcag gtcgacggta tcgataagct tgataactgg taagtttagt ctttttgtct
      961 tttatttcag gtcccggatc cggtggtggt gcaaatcaaa gaactgctcc tcagtggatg
     1021 ttgcctttac ttctaggcct gtacggaagt gttacttctg ctctaaaagc tgctctagcc
     1081 ctcgacggcg ccggcggctc ctgcaggagg ccactgtctg cagctcccgt gaagatgtcc
     1141 actccagacc cacccctggg cggaactcct cggccaggtc cttccccggg ccctggccct
     1201 tcccctggag ccatgctggg ccctagcccg ggtccctcgc cgggctccgc ccacagcatg
     1261 atggggccca gcccagggcc gccctcagca ggacacccca tccccaccca ggggcctgga
     1321 gggtaccctc aggacaacat gcaccagatg cacaagccca tggagtccat gcatgagaag
     1381 ggcatgtcgg acgacccgcg ctacaaccag atgaaaggaa tggggatgcg gtcagggggc
     1441 catgctggga tggggccccc gcccagcccc atggaccagc actcccaagg ttacccctcg
     1501 cccctgggtg gctctgagca tgcctctagt ccagttccag ccagtggccc gtcttcgggg
     1561 ccccagatgt cttccgggcc aggaggtgcc ccgctggatg gtgctgaccc ccaggccttg
     1621 gggcagcaga accggggccc aaccccattt aaccagaacc agctgcacca gctcagagct
     1681 cagatcatgg cctacaagat gctggccagg gggcagcccc tccccgacca cctgcagatg
     1741 gcggtgcagg gcaagcggcc gatgcccggg atgcagcagc agatgccaac gctacctcca
     1801 ccctcggtgt ccgcaacagg acccggccct ggccctggcc ctggccccgg cccgggtccc
     1861 ggcccggcac ctccaaatta cagcaggcct catggtatgg gagggcccaa catgcctccc
     1921 ccaggaccct cgggcgtgcc ccccgggatg ccaggccagc ctcctggagg gcctcccaag
     1981 ccctggcctg aaggacccat ggcgaatgct gctgccccca cgagcacccc tcagaagctg
     2041 attcccccgc agccaacggg ccgcccttcc cccgcgcccc ctgccgtccc acccgccgcc
     2101 tcgcccgtga tgccaccgca gacccagtcc cccgggcagc cggcccagcc cgcgcccatg
     2161 gtgccactgc accagaagca gagccgcatc acccccatcc agaagccgcg gggcctcgac
     2221 cctgtggaga tcctgcagga gcgcgagtac aggctgcagg ctcgcatcgc acaccgaatt
     2281 caggaacttg aaaaccttcc cgggtccctg gccggggatt tgcgaaccaa agcgaccatt
     2341 gagctcaagg ccctcaggct gctgaacttc cagaggcagc tgcgccagga ggtggtggtg
     2401 tgcatgcgga gggacacagc gctggagaca gccctcaatg ctaaggccta caagcgcagc
     2461 aagcgccagt ccctgcgcga ggcccgcatc actgagaagc tggagaagca gcagaagatc
     2521 gagcaggagc gcaagcgccg gcagaagcac caggaatacc tcaatagcat tctccagcat
     2581 gccaaggatt tcaaggaata tcacagatcc gtcacaggca aaatccagaa gctgaccaag
     2641 gcagtggcca cgtaccatgc caacacggag cgggagcaga agaaagagaa cgagcggatc
     2701 gagaaggagc gcatgcggag gctcatggct gaagatgagg aggggtaccg caagctcatc
     2761 gaccagaaga aggacaagcg cctggcctac ctcttgcagc agacagacga gtacgtggct
     2821 aacctcacgg agctggtgcg gcagcacaag gctgcccagg tcgccaagga gaaaaagaag
     2881 aaaaagaaaa agaagaaggc agaaaatgca gaaggacaga cgcctgccat tgggccggat
     2941 ggcgagcctc tggacgagac cagccagatg agcgacctcc cggtgaaggt gatccacgtg
     3001 gagagtggga agatcctcac aggcacagat gcccccaaag ccgggcagct ggaggcctgg
     3061 ctcgagatga acccggggta tgaagtagct ccgaggtctg atagtgaaga aagtggctca
     3121 gaagaagagg aagaggagga ggaggaagag cagccgcagg cagcacagcc tcccaccctg
     3181 cccgtggagg agaagaagaa gattccagat ccagacagcg atgacgtctc tgaggtggac
     3241 gcgcggcaca tcattgagaa tgccaagcaa gatgtcgatg atgaatatgg cgtgtcccag
     3301 gcccttgcac gtggcctgca gtcctactat gccgtggccc atgctgtcac tgagagagtg
     3361 gacaagcagt tagcgcttat ggtcaatggt gtcctcaaac agtaccagat caaaggtttg
     3421 gagtggctgg tgtccctgta caacaacaac ctgaacggca tcctggccga cgagatgggc
     3481 ctggggaaga ccatccagac catcgcgctc atcacgtacc tcatggagca caaacgcatc
     3541 aatgggccct tcctcatcat cgtgcctctc tcaacgctgt ccaactgggc gtacgagttt
     3601 gacaagtggg ccccctccgt ggtgaaggtg tcttacaagg gatccccagc agcaagacgg
     3661 gcctttgtcc cccagctccg gagtgggaag ttcaacgtct tgctgacgac gtacgagtac
     3721 atcatcaaag acaagcacat cctcgccaag atccgttgga agtacatgat tgtggacgaa
     3781 ggtcaccgca tgaagaacca ccactgcaag ctgacgcagg tgctcaacac gcactatgtg
     3841 gcaccccgcc gcctgctgct gacgggcaca ccgctgcaga acaagcttcc cgagctctgg
     3901 gcgctgctca acttcctgct gcccaccatc ttcaagagct gcagcacctt cgagcagtgg
     3961 tttaacgcac cctttgccat gaccggggaa aaggtggacc tgaatgagga ggaaaccatt
     4021 ctcatcatcc ggcgtctcca caaagtgctg cggcccttct tgctccgacg actcaagaag
     4081 gaagtcgagg cccagttgcc cgaaaaggtg gagtacgtca tcaagtgcga catgtctgcg
     4141 ctgcagcgag tgctctaccg ccacatgcag gccaagggcg tgctgctgac tgatggctcc
     4201 gagaaggaca agaagggcaa aggcggcacc aagaccctga tgaacaccat catgcagctg
     4261 cggaagatct gcaaccaccc ctacatgttc cagcacatcg aggagtcctt ttccgagcac
     4321 ttggggttca ctggcggcat tgtccaaggg ctggacctgt accgagcctc gggtaaattt
     4381 gagcttcttg atagaattct tcccaaactc cgagcaacca accacaaagt gctgctgttc
     4441 tgccaaatga cctccctcat gaccatcatg gaagattact ttgcgtatcg cggctttaaa
     4501 tacctcaggc ttgatggaac cacgaaggcg gaggaccggg gcatgctgct gaaaaccttc
     4561 aacgagcccg gctctgagta cttcatcttc ctgctcagca cccgggctgg ggggctcggc
     4621 ctgaacctcc agtcggcaga cactgtgatc atttttgaca gcgactggaa tcctcaccag
     4681 gacctgcaag cgcaggaccg agcccaccgc atcgggcagc agaacgaggt gcgtgtgctc
     4741 cgcctctgca ccgtcaacag cgtggaggag aagatcctag ctgcagccaa gtacaagctc
     4801 aacgtggacc agaaggtgat ccaggccggc atgttcgacc agaagtcctc cagccatgag
     4861 cggcgcgcct tcctgcaggc catcctggag cacgaggagc aggatgagga ggaagacgag
     4921 gtgcccgacg acgagaccgt caaccagatg atcgcccggc acgaggagga gtttgatctg
     4981 ttcatgcgca tggacctgga ccgcaggcgc gaggaggccc gcaaccccaa gcggaagccg
     5041 cgcctcatgg aggaggacga gctcccctcg tggatcatca aggacgacgc ggaggtggag
     5101 cggctgacct gtgaggagga ggaggagaag atgttcggcc gtggctcccg ccaccgcaag
     5161 gaggtggact acagcgactc actgacggag aagcagtggc tcaaggccat cgaggagggc
     5221 acgctggagg agatcgaaga ggaggtccgg cagaagaaat catcacggaa gcgcaagcga
     5281 gacagcgacg ccggctcctc caccccgacc accagcaccc gcagccgcga caaggacgac
     5341 gagagcaaga agcagaagaa gcgcgggcgg ccgcctgccg agaaactctc ccctaaccca
     5401 cccaacctca ccaagaagat gaagaagatt gtggatgccg tgatcaagta caaggacagc
     5461 agcagtggac gtcagctcag cgaggtcttc atccagctgc cctcgcgaaa ggagctgccc
     5521 gagtactacg agctcatccg caagcccgtg gacttcaaga agataaagga gcgcattcgc
     5581 aaccacaagt accgcagcct caacgaccta gagaaggacg tcatgctcct gtgccagaac
     5641 gcacagacct tcaacctgga gggctccctg atctatgaag actccatcgt cttgcagtcg
     5701 gtcttcacca gcgtgcggca gaaaatcgag aaggaggatg acagtgaagg cgaggagagt
     5761 gaggaggagg aagagggcga ggaggaaggc tccgaatccg aatctcggtc cgtcaaagtg
     5821 aagatcaagc ttggccggaa ggagaaggca caggaccggc tgaagggcgg ccggcggcgg
     5881 ccgagccgag ggtcccgagc caagccggtc gtgagtgacg atgacagtga ggaggaacaa
     5941 gaggaggacc gctcaggaag tggcagcgaa gaagacgact acaaggacga cgatgacaaa
     6001 tgataagtcg actctagagg atcccgggtg gcatccctgt gacccctccc cagtgcctct
     6061 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     6121 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     6181 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     6241 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     6301 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     6361 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     6421 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     6481 cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca
     6541 gacacagcat aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg
     6601 cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc
     6661 agcttggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     6721 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     6781 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     6841 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6901 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6961 ttttggaggc ctaggctttt gcaaaaagct cctcgaggaa ctgaaaaacc agaaagttaa
     7021 ttccctatag tgagtcgtat taaattcgta atcatgtcat agctgtttcc tgtgtgaaat
     7081 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     7141 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     7201 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7261 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     7321 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7381 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7441 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7501 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7561 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7621 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7681 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7741 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7801 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7861 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7921 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7981 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     8041 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     8101 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     8161 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     8221 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     8281 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     8341 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     8401 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8461 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8521 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8581 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8641 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8701 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8761 actggtgagt actcaaccaa gtcattctga gaatagtata tgcggcgacc gagttgctct
     8821 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8881 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8941 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     9001 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     9061 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     9121 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     9181 cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg
     9241 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     9301 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     9361 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     9421 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     9481 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     9541 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     9601 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     9661 atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
//
