LOCUS       sequence-169373-        5806 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 5806)
  AUTHORS   Hajra KM, Ji X, Fearon ER
  TITLE     Extinction of E-cadherin expression in breast cancer via a dominant 
            repression pathway acting on proximal promoter elements.
  JOURNAL   Oncogene. 1999 Dec 2. 18(51):7274-9.
  PUBMED    10602481
REFERENCE   2  (bases 1 to 5806)
  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: "Oncogene. 
            1999 Dec 2. 18(51):7274-9."
COMMENT     Sequence Label: pGL2Basic-EcadK1
FEATURES             Location/Qualifiers
     source          1..5806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             431..2083
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(497..515)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          2324..2389
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2519..2539
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2964..3098
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3240..3257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3411..3999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3491..3510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4170..4961,4962..5030))
                     /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     4793..4812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5031..5135)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5162..5617
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5249..5268)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5459..5480
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
       61 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
      121 tatggtactg taactgagct aacataaccc gggaggtacc gagctcgtcc gcgctgctga
      181 ttggctgtgg ccggcaggtg aaccctcagc caatcagcgg tacggggggc ggtgcctccg
      241 gggctcacct ggctgcagcc acgcaccccc tctcagtggc gtcggaactg caaagcacct
      301 gtgagcttgc ggaagtcagt tcagactcca gcccgctcca gcccggcccg acccgaccgc
      361 acccggcgcc tgccctcgct cggcgtcccc ggcagccacc caagcttggc attccggtac
      421 tgttggtaaa atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatcctct
      481 agaggatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc
      541 tggaacaatt gcttttacag atgcacatat cgaggtgaac atcacgtacg cggaatactt
      601 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag
      661 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt
      721 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag
      781 tatgaacatt tcgcagccta ccgtagtgtt tgtttccaaa aaggggttgc aaaaaatttt
      841 gaacgtgcaa aaaaaattac caataatcca gaaaattatt atcatggatt ctaaaacgga
      901 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa
      961 tgaatacgat tttgtaccag agtcctttga tcgtgacaaa acaattgcac tgataatgaa
     1021 ttcctctgga tctactgggt tacctaaggg tgtggccctt ccgcatagaa ctgcctgcgt
     1081 cagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat
     1141 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat
     1201 atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttt tacgatccct
     1261 tcaggattac aaaattcaaa gtgcgttgct agtaccaacc ctattttcat tcttcgccaa
     1321 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctgggggcgc
     1381 acctctttcg aaagaagtcg gggaagcggt tgcaaaacgc ttccatcttc cagggatacg
     1441 acaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga
     1501 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga
     1561 taccgggaaa acgctgggcg ttaatcagag aggcgaatta tgtgtcagag gacctatgat
     1621 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg
     1681 gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca tagttgaccg
     1741 cttgaagtct ttaattaaat acaaaggata tcaggtggcc cccgctgaat tggaatcgat
     1801 attgttacaa caccccaaca tcttcgacgc gggcgtggca ggtcttcccg acgatgacgc
     1861 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga
     1921 gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt
     1981 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga
     2041 gatcctcata aaggccaaga agggcggaaa gtccaaattg taaaatgtaa ctgtattcag
     2101 cgatgacgaa attcttagct attgtaatac tgcgatgagt ggcagggcgg ggcgtaattt
     2161 ttttaaggca gttattggtg cccttaaacg cctggttgct acgcctgaat aagtgataat
     2221 aagcggatga atggcagaaa ttcgccggat ctttgtgaag gaaccttact tctgtggtgt
     2281 gacataattg gacaaactac ctacagagat ttaaagctct aaggtaaata taaaattttt
     2341 aagtgtataa tgtgttaaac tactgattct aattgtttgt gtattttaga ttccaaccta
     2401 tggaactgat gaatgggagc agtggtggaa tgcctttaat gaggaaaacc tgttttgctc
     2461 agaagaaatg ccatctagtg atgatgaggc tactgctgac tctcaacatt ctactcctcc
     2521 aaaaaagaag agaaaggtag aagaccccaa ggactttcct tcagaattgc taagtttttt
     2581 gagtcatgct gtgtttagta atagaactct tgcttgcttt gctatttaca ccacaaagga
     2641 aaaagctgca ctgctataca agaaaattat ggaaaaatat tctgtaacct ttataagtag
     2701 gcataacagt tataatcata acatactgtt ttttcttact ccacacaggc atagagtgtc
     2761 tgctattaat aactatgctc aaaaattgtg tacctttagc tttttaattt gtaaaggggt
     2821 taataaggaa tatttgatgt atagtgcctt gactagagat cataatcagc cataccacat
     2881 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     2941 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3001 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3061 tgtccaaact catcaatgta tcttatcatg tctggatccg tcgaccgatg cccttgagag
     3121 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     3181 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc
     3241 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3301 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3361 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3421 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3481 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3541 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3601 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3661 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3721 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3781 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3841 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3901 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3961 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4021 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4081 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4141 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4201 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4261 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4321 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4381 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4441 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4501 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4561 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4621 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4681 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4741 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4801 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4861 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4921 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4981 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5041 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5101 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5161 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     5221 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     5281 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     5341 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     5401 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5461 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5521 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5581 aacgcgaatt ttaacaaaat attaacgctt acaatttgcc attcgccatt caggctgcgc
     5641 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc caagctacca
     5701 tgataagtaa gtaatattaa ggtacgtgga ggttttactt gctttaaaaa acctcccaca
     5761 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgtt
//
