LOCUS       sequence-424665-        6628 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 6628)
  AUTHORS   Lee K, Gjorevski N, Boghaert E, Radisky DC, Nelson CM
  TITLE     Snail1, Snail2, and E47 promote mammary epithelial branching 
            morphogenesis.
  JOURNAL   EMBO J. 2011 May 24;30(13):2662-74. doi: 10.1038/emboj.2011.159.
  PUBMED    21610693
REFERENCE   2  (bases 1 to 6628)
  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"; doi: 
            "10.1038/emboj.2011"; journalName: "EMBO J"; date: "2011-05-24- 24";
            volume: "30"; issue: "13"; pages: "2662-74"
COMMENT     Sequence Label: p3xFLAG-mE47
FEATURES             Location/Qualifiers
     source          1..6628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(151..170)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(152..170)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(184..200)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(184..200)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(197..219)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    208..224
                     /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(232..238,239..256,257..262))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    277..298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(415..432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(586..1174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(666..685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1345..2136,2137..2205))
                     /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     1968..1987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2206..2310)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2337..2792
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2424..2443)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2634..2655
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2918..2940
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2932..2949
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2933..2949
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3110..3489
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3490..3693
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3644..3664
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3723..3788
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    5761..6383
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5960..5979)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6407..6427)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6412..6628,1..113)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6592..6628,1..99)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
       61 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
      121 tcgaggaact gaaaaaccag aaagttaatt ccctatagtg agtcgtatta aattcgtaat
      181 catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
      241 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
      301 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
      361 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
      421 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      481 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
      541 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      601 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      661 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      721 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      781 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      841 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      901 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      961 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1021 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1081 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1141 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1201 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1261 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1321 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1381 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1441 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1501 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1561 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1621 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     1681 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1741 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1801 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1861 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1921 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     1981 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2041 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2101 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2161 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2221 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2281 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
     2341 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2401 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2461 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2521 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2581 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2641 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2701 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2761 gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct gcgcaactgt
     2821 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     2881 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     2941 acggccagtg ccaagctgat ctatacattg aatcaatatt ggcaattagc catattagtc
     3001 attggttata tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat
     3061 cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgttg acattgatta
     3121 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     3181 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     3241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     3301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     3361 atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     3421 cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     3481 attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg gtttgactca
     3541 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     3601 caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg
     3661 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gaattaacca
     3721 tggactacaa agaccatgac ggtgattata aagatcatga catcgattac aaggatgacg
     3781 atgacaagct tgcggccgcg aattcaatga tgaaccagtc tcagagaatg gcacccgtgg
     3841 gctctgacaa ggaactgagt gacctcctgg acttcagcat gatgttcccg ctacctgtgg
     3901 ccaatgggaa gagccggccc gcctccctcg ggggaaccca gtttgcaggc tcaggactgg
     3961 aggaccgacc cagctcaggc tcctggggca gcagtgacca gaacagttct tcctttgacc
     4021 ctagccggac atacagcgaa ggtgcccact tcagtgactc ccacagcagc ctgccgcctt
     4081 ccacgttcct aggagctggg cttggaggca agggcagtga gcggaatgcc tatgccacct
     4141 ttgggagaga caccagtgtt ggcaccttga gtcaggctgg cttcctgcca ggtgagctga
     4201 gcctcagcag tcccgggcca ctgtccccat cgggcatcaa gagcagctcc cagtattacc
     4261 cctcattccc cagcaaccct cgtcggagag ctgcagatgg tggcctggat actcagccga
     4321 agaaggtccg gaaggttccg cctggtctcc cttcctcggt gtatccgccc agctcaggtg
     4381 acagctacag cagggatgct gcagcctacc cctccgccaa gacccccagc agcgcttacc
     4441 cctccccctt ctacgtggca gatggcagcc tgcacccatc agctgagctc tggagtacgc
     4501 ctagccaggt gggctttggg cccatgctag gtgacggctc ttcccctctg ccccttgcac
     4561 cgggcagcag ctccgtgggc agtggtacct ttgggggcct ccagcagcag gatcgcatgg
     4621 gctaccagct gcatggatct gaggttaatg gctcgctccc agctgtatcc agcttttcgg
     4681 ctgcccctgg cacttacagt gggacttccg gccacacgcc ccctgtgagt ggggccgcag
     4741 ctgaaagcct cctaggcacc cgagggacta cagccagcag ctcaggggat gcccttggga
     4801 aggcactggc ctcgatctac tccccggatc actccagcaa taatttctca cctagcccct
     4861 caacgcctgt gggttcaccc cagggcctgc cagggacatc acagtggccc cgggcaggag
     4921 cgcccagtgc cttatccccc aactacgatg caggtctcca tggcctgagc aagatggagg
     4981 accgcttgga cgaggccatc catgtcctgc gaagccacgc tgttggcacc gctagcgatc
     5041 tccatgggct tttgcctggc catggcgcac tgaccacgag cttcaccggc cccatgtcac
     5101 tgggcgggcg gcatgccggc ctggtcgggg gaagccatcc tgaggagggc ctcacaagtg
     5161 gggccagtct tttgcataac catgccagcc tccccagcca gcccagttcc ctccctgacc
     5221 tctcacagag acctcccgac tcctatagtg gactcgggag ggcaggcaca acagcgggtg
     5281 ccagcgagat caagcgggag gagaaagagg atgaggaaat cgcatcagta gccgacgccg
     5341 aagaggacaa gaaggacctg aaggtcccac gcacgcgcac cagcagtaca gatgaggtgc
     5401 tgtccctgga ggagaaggac ctgagggacc gggagaggcg tatggccaat aacgctcggg
     5461 agcgggtgcg cgtgcgggac attaacgagg ccttccggga gctgggccgc atgtgccagc
     5521 tgcacctcaa gtcggataag gcgcagacca agctgctcat cctgcagcag gcggtgcagg
     5581 tcatcctggg cctggagcag caggtgcgag aacgcaacct gaaccccaaa gcagcctgct
     5641 tgaagcggag ggaggaggag aaggtgtctg gcgtggtcgg ggacccacag ctgcccctgt
     5701 cagccgccca cccgggcctg ggtgaggccc acaacccagc cgggcacctg tgaggatccc
     5761 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     5821 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     5881 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     5941 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     6001 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     6061 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     6121 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     6181 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     6241 attttaaaat aactatacca gcaggaggac gtccagacac agcataggct acctggccat
     6301 gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc
     6361 actcagtaga tgcctgttga attgggtacg cggccagctt ggctgtggaa tgtgtgtcag
     6421 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6481 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     6541 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     6601 ctaactccgc ccagttccgc ccattctc
//
