LOCUS       Exported                5927 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    -962 human cyclin D1 promoter pGL3Basic
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5927)
  AUTHORS   Tetsu O, McCormick F
  TITLE     Beta-catenin regulates expression of cyclin D1 in colon carcinoma 
            cells.
  JOURNAL   Nature. 1999 Apr 1;398(6726):422-6.
  PUBMED    10201372
REFERENCE   2  (bases 1 to 5927)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5927
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      131..586
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(218..237)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     428..449
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    717..765
                     /note="synthetic polyadenylation signal"
     misc_feature    779..870
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     819..838
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      2068..2077
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2074..3726
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(2140..2158)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3767..3888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3778..3797)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3832..3851
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4136..4153)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4307..4895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4387..4406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5066..5926)
                     /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     5689..5708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5927..104)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
       61 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
      121 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
      181 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
      241 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
      301 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
      361 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
      421 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
      481 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
      541 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc
      661 aagctaccat gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata
      721 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa
      781 catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca
      841 gtgcaagtgc aggtgccaga acatttctct atcgataggt accgagctct tacgcgtgct
      901 agcccgggct cgagatcccc gggtactaaa aaaaatgagt cagaatggag atcactgttt
      961 ctcagctttc cattcagagg tgtgtttctc ccggttaaat tgccggcacg ggaagggagg
     1021 gggtgcagtt ggggaccccc gcaaggaccg actggtcaag gtaggaaggc agcccgaaga
     1081 gtctccaggc tagaaggaca agatgaagga aatgctggcc accatcttgg gctgctgctg
     1141 gaattttcgg gcatttattt tattttattt tttgagcgag cgcatgctaa gctgaaatcc
     1201 ctttaacttt tagggttacc cccttgggca tttgcaacga cgcccctgtg cgccggaatg
     1261 aaacttgcac aggggttgtg tgcccggtcc tccccgtcct tgcatgctaa attagttctt
     1321 gcaatttaca cgtgttaatg aaaatgaaag aagatgcagt cgctgagatt ctttggccgt
     1381 ctgtccgccc gtgggtgccc tcgtggcgtt cttggaaatg cgcccattct gccggcttgg
     1441 atatggggtg tcgccgcgcc ccagtcaccc cttctcgtgg tctccccagg ctgcgtgtgg
     1501 cctgccggcc ttcctagttg tcccctactg cagagccacc tccacctcac cccctaaatc
     1561 ccgggggacc cactcgaggc ggacggggcc ccctgcaccc ctcttccctg gcggggagaa
     1621 aggctgcagc ggggcgattt gcatttctat gaaaaccgga ctacaggggc aactccgccg
     1681 cagggcaggc gcggcgcctc agggatggct tttgggctct gcccctcgct gctcccggcg
     1741 tttggcgccc gcgccccctc cccctgcgcc cgcccccgcc cccctcccgc tcccattctc
     1801 tgccgggctt tgatctttgc ttaacaacag taacgtcaca cggactacag gggagttttg
     1861 ttgaagttgc aaagtcctgg agcctccaga gggctgtcgg cgcagtagca gcgagcagca
     1921 gagtccgcac gctccggcga ggggcagaag agcgcgaggg agcgcggggc agcagaagcg
     1981 agagccgagc gcggacccag ccaggaccca cagccctccc caggggttcg aaatcgataa
     2041 gcttggcatt ccggtactgt tggtaaagcc accatggaag acgccaaaaa cataaagaaa
     2101 ggcccggcgc cattctatcc gctggaagat ggaaccgctg gagagcaact gcataaggct
     2161 atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca tatcgaggtg
     2221 gacatcactt acgctgagta cttcgaaatg tccgttcggt tggcagaagc tatgaaacga
     2281 tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct tcaattcttt
     2341 atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa cgacatttat
     2401 aatgaacgtg aattgctcaa cagtatgggc atttcgcagc ctaccgtggt gttcgtttcc
     2461 aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat ccaaaaaatt
     2521 attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac gttcgtcaca
     2581 tctcatctac ctcccggttt taatgaatac gattttgtgc cagagtcctt cgatagggac
     2641 aagacaattg cactgatcat gaactcctct ggatctactg gtctgcctaa aggtgtcgct
     2701 ctgcctcata gaactgcctg cgtgagattc tcgcatgcca gagatcctat ttttggcaat
     2761 caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg ttttggaatg
     2821 tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta tagatttgaa
     2881 gaagagctgt ttctgaggag ccttcaggat tacaagattc aaagtgcgct gctggtgcca
     2941 accctattct ccttcttcgc caaaagcact ctgattgaca aatacgattt atctaattta
     3001 cacgaaattg cttctggtgg cgctcccctc tctaaggaag tcggggaagc ggttgccaag
     3061 aggttccatc tgccaggtat caggcaagga tatgggctca ctgagactac atcagctatt
     3121 ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt
     3181 gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca aagaggcgaa
     3241 ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa acaatccgga agcgaccaac
     3301 gccttgattg acaaggatgg atggctacat tctggagaca tagcttactg ggacgaagac
     3361 gaacacttct tcatcgttga ccgcctgaag tctctgatta agtacaaagg ctatcaggtg
     3421 gctcccgctg aattggaatc catcttgctc caacacccca acatcttcga cgcaggtgtc
     3481 gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt tttggagcac
     3541 ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg
     3601 aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa
     3661 ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggcgg aaagatcgcc
     3721 gtgtaattct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat
     3781 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3841 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3901 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     3961 aacctctaca aatgtggtaa aatcgataag gatccgtcga ccgatgccct tgagagcctt
     4021 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     4081 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     4141 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4201 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4261 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4321 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4381 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4441 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4501 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4561 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4621 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4681 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4741 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4801 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4861 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4921 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4981 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5041 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5101 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5161 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5221 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5281 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5341 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5401 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5461 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5521 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5581 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5641 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5701 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5761 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5821 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5881 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcata
//
