LOCUS       sequence-140267-        5108 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 5108)
  AUTHORS   Gomis RR, Alarcon C, Nadal C, Van Poznak C, Massague J
  TITLE     C/EBPbeta at the core of the TGFbeta cytostatic response and its 
            evasion in metastatic breast cancer cells.
  JOURNAL   Cancer Cell. 2006 Sep . 10(3):203-14.
  PUBMED    16959612
REFERENCE   2  (bases 1 to 5108)
  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: "Cancer 
            Cell. 2006 Sep . 10(3):203-14."
COMMENT     Sequence Label: pCMV-FLAG LIP
FEATURES             Location/Qualifiers
     source          1..5108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1985..2440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2072..2091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2282..2303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2566..2588
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2580..2597
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2581..2597
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2758..3137
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3138..3341
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3292..3312
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3376..3399
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3910..4532
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4109..4128)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4556..4576)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4721..4855
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4907..4926)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4908..4926)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4940..4956)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4940..4956)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4953..4975)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4964..4980
                     /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(4988..4994,4995..5012,5013..5018))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5033..5054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2341 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2401 tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc
     2461 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     2521 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     2581 gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca
     2641 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
     2701 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
     2761 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     2821 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2881 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2941 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     3001 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     3061 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
     3121 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
     3181 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     3241 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
     3301 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     3361 attgcagtga ccatggacta caaggacgac gatgacaagg aattcagcat ggcggcgggc
     3421 ttcccgtacg cgctgcgcgc ttacctcggc taccaggcgg tgccgagcgg cagcagcggg
     3481 agcctctcca cgtcctcctc gtccagcccg cccggcacgc cgagccccgc tgacgccaag
     3541 gcgcccccga ccgcctgcta cgcgggggcc gcgccggcgc cctcgcaggt caagagcaag
     3601 gccaagaaga ccgtggacaa gcacagcgac gagtacaaga tccggcgcga gcgcaacaac
     3661 atcgccgtgc gcaagagccg cgacaaggcc aagatgcgca acctggagac gcagcacaag
     3721 gtcctggagc tcacggccga gaacgagcgg ctgcagaaga aggtggagca gctgtcgcgc
     3781 gagctcagca ccctgcggaa cttgttcaag cagctgcccg agcccctgct cgcctcctcc
     3841 ggccactgct agcgggtacc acgcgtatcg ataagcttgc atgcctgcag gtcgactcta
     3901 gaggatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     3961 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     4021 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     4081 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     4141 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     4201 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     4261 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     4321 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     4381 gtccttctga ttttaaaata actataccag caggaggacg tccagacaca gcataggcta
     4441 cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt cctctcatgc
     4501 gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcttg gctgtggaat
     4561 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4621 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4681 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4741 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4801 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4861 ttttgcaaaa agctcctcga ggaactgaaa aaccagaaag ttaattccct atagtgagtc
     4921 gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4981 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5041 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5101 gccagctg
//
