LOCUS       sequence-424324-        5304 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 5304)
  AUTHORS   Yu J, Zhang L, Hwang PM, Kinzler KW, Vogelstein B
  TITLE     PUMA induces the rapid apoptosis of colorectal cancer cells.
  JOURNAL   Mol Cell. 2001 Mar . 7(3):673-82.
  PUBMED    11463391
REFERENCE   2  (bases 1 to 5304)
  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: "Mol Cell. 
            2001 Mar . 7(3):673-82."
COMMENT     Sequence Label: PUMA Frag1-Luc
FEATURES             Location/Qualifiers
     source          1..5304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    7..55
                     /note="synthetic polyadenylation signal"
     misc_feature    69..160
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     109..128
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      735..744
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             741..2393
                     /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(807..825)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2434..2555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2445..2464)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2499..2518
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2803..2820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2974..3562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3054..3073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3733..4524,4525..4593))
                     /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     4356..4375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4594..4698)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4725..5180
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4812..4831)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5022..5043
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
       61 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
      121 gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgataggt accgcgagac
      181 tgtggccttg tgtctgtgag tacatcctct gggctctgcc tgcacgtgac tttgtggacc
      241 ctggaacgcc cgtcggtcgg tctgtgtacg catcgctggg ggtgtggatc tgtgggtccc
      301 agtcagtgtg tgtgtccgac tgtcccggtg tctgggcgat ctccccacac cccgccgcac
      361 agcgcctggg tcctccttgc cttgggctag gccctgcccg tcccccgctg cagggaaacc
      421 cccggcgcgg aggcgggggg ggcgcggcgc gcgcctgcaa gtcctgactt gtccgcggcg
      481 ggcggggccg tagcgtcacg cgggggcggg gcgtgggacc cgccgggcgg gggcggggcg
      541 gggcggggcg gggcggcttt ggagcgggcc cgggatccgc cgggcggcat gagacgcggc
      601 gcgagccaca tgcgagcggg cgcctggcgg cggcggcggc accagcgatc ccagcagcga
      661 attcgggtat ataatggatc cggtatcgag atctgcgatc taagtaagct tggcattccg
      721 gtactgttgg taaagccacc atggaagacg ccaaaaacat aaagaaaggc ccggcgccat
      781 tctatccgct ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg
      841 ccctggttcc tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg
      901 ctgagtactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata
      961 caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg
     1021 gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat
     1081 tgctcaacag tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc
     1141 aaaaaatttt gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt
     1201 ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc
     1261 ccggttttaa tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac
     1321 tgatcatgaa ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa
     1381 ctgcctgcgt gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg
     1441 atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg
     1501 gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttc
     1561 tgaggagcct tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct
     1621 tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt
     1681 ctggtggcgc tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc
     1741 caggtatcag gcaaggatat gggctcactg agactacatc agctattctg attacacccg
     1801 agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg
     1861 tggatctgga taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag
     1921 gtcctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca
     1981 aggatggatg gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca
     2041 tcgttgaccg cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat
     2101 tggaatccat cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg
     2161 acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga
     2221 cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg
     2281 gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa
     2341 aaatcagaga gatcctcata aaggccaaga agggcggaaa gatcgccgtg taattctaga
     2401 gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     2461 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     2521 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     2581 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     2641 gtggtaaaat cgataaggat ccgtcgaccg atgcccttga gagccttcaa cccagtcagc
     2701 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
     2761 atgcaactcg taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct
     2821 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2881 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2941 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3001 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3061 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3121 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3181 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3241 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3301 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3361 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3421 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3481 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3541 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3601 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3661 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3721 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3781 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3841 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3901 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3961 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4021 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4081 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4141 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4201 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4261 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4321 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4381 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4441 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4501 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4561 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4621 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4681 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     4741 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4801 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4861 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4921 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4981 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5041 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     5101 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5161 aatattaacg cttacaattt gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     5221 atcggtgcgg gcctcttcgc tattacgcca gcccaagcta ccatgataag taagtaatat
     5281 taaggtacgg gaggtacttg gagc
//
