LOCUS       sequence-352774-        4389 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4389)
  AUTHORS   St-Pierre F, Cui L, Priest DG, Endy D, Dodd IB, Shearwin KE
  TITLE     One-step cloning and chromosomal integration of DNA.
  JOURNAL   ACS Synth Biol. 2013 Sep 20;2(9):537-41. doi: 10.1021/sb400021j. 
            Epub 2013 May 20.
  PUBMED    24050148
REFERENCE   2  (bases 1 to 4389)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1021/sb400021j";
            journalName: "ACS Synth Biol"; date: "2013-09-20- 20"; volume: "2"; 
            issue: "9"; pages: "537-41"
COMMENT     Sequence Label: pE-FLP
FEATURES             Location/Qualifiers
     source          1..4389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(286..1077,1078..1146))
                     /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     909..928
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1147..1251)
                     /gene="bla"
                     /label=AmpR promoter
     RBS             1353..1361
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1365..2636
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=FLP
                     /note="FLP is a site-specific recombinase from 
                     Saccharomyces cerevisiae. Recombination occurs at FRT 
                     sequences."
                     /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     CDS             complement(2833..3783)
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3831..4053
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ctgctatgtg gtgctatctg actttttgct gttcagcagt tcctgccctc tgattttcca
       61 gtctgaccac ttcggattat cccgtgacag gtcattcaga ctggctaatg cacccagtaa
      121 ggcagcggta tcatcaacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      181 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
      241 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
      301 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
      361 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
      421 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
      481 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
      541 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
      601 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
      661 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
      721 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
      781 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
      841 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
      901 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
      961 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1021 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1081 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1141 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1201 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     1261 ccgaaaagtg ccacctgctt atcgtgtttg acatggtgtt tagatctcaa tagtatttag
     1321 tttagatgta gattgtttag tgcttgatgt actaaggagg ttgtatgcca caatttgata
     1381 tattatgtaa aacaccacct aaggtgcttg ttcgtcagtt tgtggaaagg tttgaaagac
     1441 cttcaggtga gaaaatagca ttatgtgctg ctgaactaac ctatttatgt tggatgatta
     1501 cacataacgg aacagcaatc aagagagcca cattcatgag ctataatact atcataagca
     1561 attcgctgag tttcgatatt gtcaataaat cactccagtt taaatacaag acgcaaaaag
     1621 caacaattct ggaagcctca ttaaagaaat tgattcctgc ttgggaattt acaattattc
     1681 cttactatgg acaaaaacat caatctgata tcactgatat tgtaagtagt ttgcaattac
     1741 agttcgaatc atcggaagaa gcagataagg gaaatagcca cagtaaaaaa atgcttaaag
     1801 cacttctaag tgagggtgaa agcatctggg agatcactga gaaaatacta aattcgtttg
     1861 agtatacttc gagatttaca aaaacaaaaa ctttatacca attcctcttc ctagctactt
     1921 tcatcaattg tggaagattc agcgatatta agaacgttga tccgaaatca tttaaattag
     1981 tccaaaataa gtatctggga gtaataatcc agtgtttagt gacagagaca aagacaagcg
     2041 ttagtaggca catatacttc tttagcgcaa ggggtaggat cgatccactt gtatatttgg
     2101 atgaattttt gaggaattct gaaccagtcc taaaacgagt aaataggacc ggcaattctt
     2161 caagcaataa acaggaatac caattattaa aagataactt agtcagatcg tacaataaag
     2221 ctttgaagaa aaatgcgcct tattcaatct ttgctataaa aaatggccca aaatctcaca
     2281 ttggaagaca tttgatgacc tcatttcttt caatgaaggg cctaacggag ttgactaatg
     2341 ttgtgggaaa ttggagcgat aagcgtgctt ctgccgtggc caggacaacg tatactcatc
     2401 agataacagc aatacctgat cactacttcg cactagtttc tcggtactat gcatatgatc
     2461 caatatcaaa ggaaatgata gcattgaagg atgagactaa tccaattgag gagtggcagc
     2521 atatagaaca gctaaagggt agtgctgaag gaagcatacg ataccccgca tggaatggga
     2581 taatatcaca ggaggtacta gactaccttt catcctacat aaatagacgc atataaccat
     2641 ggcatgcatg gtatcgagat ggcacatagc cttgctcaaa ttggaatcag gtttgtgcca
     2701 ataccagtag aaacagacga agaatccatg ggtatggaca gttttccctt tgatatgtaa
     2761 cggtgaacag ttgttctact tttgtttgtt agtcttgatg cttcactgat agatacaaga
     2821 gccataagaa cctcagatcc ttccgtattt agccagtatg ttctctagtg tggttcgttg
     2881 tttttgcgtg agccatgaga acgaaccatt gagatcatac ttactttgca tgtcactcaa
     2941 aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt
     3001 tcttagtccg ttatgtaggt aggaatctga tgtaatggtt gttggtattt tgtcaccatt
     3061 catttttatc tggttgttct caagttcggt tacgagatcc atttgtctat ctagttcaac
     3121 ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca accaccaatt tcatattgct
     3181 gtaagtgttt aaatctttac ttattggttt caaaacccat tggttaagcc ttttaaactc
     3241 atggtagtta ttttcaagca ttaacatgaa cttaaattca tcaaggctaa tctctatatt
     3301 tgccttgtga gttttctttt gtgttagttc ttttaataac cactcataaa tcctcataga
     3361 gtatttgttt tcaaaagact taacatgttc cagattatat tttatgaatt tttttaactg
     3421 gaaaagataa ggcaatatct cttcactaaa aactaattct aatttttcgc ttgagaactt
     3481 ggcatagttt gtccactgga aaatctcaaa gcctttaacc aaaggattcc tgatttccac
     3541 agttctcgtc atcagctctc tggttgcttt agctaataca ccataagcat tttccctact
     3601 gatgttcatc atctgaacgt attggttata agtgaacgat accgtccgtt ctttccttgt
     3661 agggttttca atcgtggggt tgagtagtgc cacacagcat aaaattagct tggtttcatg
     3721 ctccgttaag tcatagcgac taatcgctag ttcatttgct ttgaaaacaa ctaattcaga
     3781 catacatctc aattggtcta ggtgatttta atcactatac caattgagat gggctagtca
     3841 atgataatta ctagtccttt tcctttgagt tgtgggtatc tgtaaattct gctagacctt
     3901 tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt
     3961 tttttttgtt tatattcaag tggttataat ttatagaata aagaaagaat aaaaaaagat
     4021 aaaaagaata gatcccagcc ctgtgtataa ctcactactt tagtcagttc cgcagtatta
     4081 caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa accctaaagg
     4141 cttaagtagc accctcgcaa gctcggttgc ggccgcaatc gggcaaatcg ctgaatattc
     4201 cttttgtctc cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat tcagttcgct
     4261 gcgctcacgg ctctggcagt gaatgggggt aaatggcact acaggcgcct tttatggatt
     4321 catgcaagga aactacccat aatacaagaa aagcccgtca cgggcttctc agggcgtttt
     4381 atggcgggt
//
