LOCUS       sequence-204984-        5633 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 5633)
  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 5633)
  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 LAP2
FEATURES             Location/Qualifiers
     source          1..5633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        59..438
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        439..642
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     593..613
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             677..700
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1736..2358
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1935..1954)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2382..2402)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2547..2681
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2733..2752)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2734..2752)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2766..2782)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2766..2782)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2779..2801)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2790..2806
                     /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(2814..2820,2821..2838,2839..2844))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2859..2880
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2997..3014)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3168..3756)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3248..3267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3927..4718,4719..4787))
                     /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     4550..4569
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4788..4892)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4919..5374
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5006..5025)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5216..5237
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5500..5522
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5514..5531
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5515..5531
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc gccatgttga
       61 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      121 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      181 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      241 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      301 gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      361 cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat ctacgtatta
      421 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg
      481 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      541 caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg
      601 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
      661 aattgcagtg accatggact acaaggacga cgatgacaag gaattcacca tggaagtggc
      721 caacttctac tacgaggcgg actgcttggc tgctgcgtac ggcggcaagg cggcccccgc
      781 ggcgcccccc gcggccagac ccgggccgcg cccccccgcc ggcgagctgg gcagcatcgg
      841 cgaccacgag cgcgccatcg acttcagccc gtacctggag ccgctgggcg cgccgcaggc
      901 cccggcgccc gccacggcca cggacacctt cgaggcggct ccgcccgcgc ccgcccccgc
      961 gcccgcctcc tccgggcagc accacgactt cctctccgac ctcttctccg acgactacgg
     1021 gggcaagaac tgcaagaagc cggccgagta cggctacgtg agcctggggc gcctgggggc
     1081 tgccaagggc gcgctgcacc ccggctgctt cgcgcccctg cacccaccgc ccccgccgcc
     1141 gccgccgccc gccgagctca aggcggagcc gggcttcgag cccgcggact gcaagcggaa
     1201 ggaggaggcc ggggcgccgg gcggcggcgc aggcatggcg gcgggcttcc cgtacgcgct
     1261 gcgcgcttac ctcggctacc aggcggtgcc gagcggcagc agcgggagcc tctccacgtc
     1321 ctcctcgtcc agcccgcccg gcacgccgag ccccgctgac gccaaggcgc ccccgaccgc
     1381 ctgctacgcg ggggccgcgc cggcgccctc gcaggtcaag agcaaggcca agaagaccgt
     1441 ggacaagcac agcgacgagt acaagatccg gcgcgagcgc aacaacatcg ccgtgcgcaa
     1501 gagccgcgac aaggccaaga tgcgcaacct ggagacgcag cacaaggtcc tggagctcac
     1561 ggccgagaac gagcggctgc agaagaaggt ggagcagctg tcgcgcgagc tcagcaccct
     1621 gcggaacttg ttcaagcagc tgcccgagcc cctgctcgcc tcctccggcc actgctagcg
     1681 ggtaccacgc gtatcgataa gcttgcatgc ctgcaggtcg actctagagg atcccgggtg
     1741 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     1801 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     1861 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     1921 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     1981 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     2041 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     2101 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     2161 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     2221 aaaataacta taccagcagg aggacgtcca gacacagcat aggctacctg gccatgccca
     2281 accggtggga catttgagtt gcttgcttgg cactgtcctc tcatgcgttg ggtccactca
     2341 gtagatgcct gttgaattgg gtacgcggcc agcttggctg tggaatgtgt gtcagttagg
     2401 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2461 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2521 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccagtt ccgcccattc
     2581 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     2641 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     2701 cctcgaggaa ctgaaaaacc agaaagttaa ttccctatag tgagtcgtat taaattcgta
     2761 atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2821 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2881 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2941 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3001 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3061 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3121 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3181 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3241 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3301 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3361 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3421 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3481 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3541 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3601 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3661 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3721 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3781 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3841 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3901 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3961 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4021 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4081 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4141 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4201 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4261 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4321 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4381 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4441 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4501 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4561 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4621 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4681 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4741 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4801 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4861 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4921 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4981 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5041 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5101 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5161 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5221 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5281 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5341 gcgaatttta acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact
     5401 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     5461 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     5521 cgacggccag tgccaagctg atctatacat tgaatcaata ttggcaatta gccatattag
     5581 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttg
//
