LOCUS       Exported                4389 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pE-FLP
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 Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4389
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     RBS             76..84
                     /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             88..1359
                     /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(1556..2506)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2554..2776
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(3398..4258)
                     /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     4021..4040
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4259..4363)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cttatcgtgt ttgacatggt gtttagatct caatagtatt tagtttagat gtagattgtt
       61 tagtgcttga tgtactaagg aggttgtatg ccacaatttg atatattatg taaaacacca
      121 cctaaggtgc ttgttcgtca gtttgtggaa aggtttgaaa gaccttcagg tgagaaaata
      181 gcattatgtg ctgctgaact aacctattta tgttggatga ttacacataa cggaacagca
      241 atcaagagag ccacattcat gagctataat actatcataa gcaattcgct gagtttcgat
      301 attgtcaata aatcactcca gtttaaatac aagacgcaaa aagcaacaat tctggaagcc
      361 tcattaaaga aattgattcc tgcttgggaa tttacaatta ttccttacta tggacaaaaa
      421 catcaatctg atatcactga tattgtaagt agtttgcaat tacagttcga atcatcggaa
      481 gaagcagata agggaaatag ccacagtaaa aaaatgctta aagcacttct aagtgagggt
      541 gaaagcatct gggagatcac tgagaaaata ctaaattcgt ttgagtatac ttcgagattt
      601 acaaaaacaa aaactttata ccaattcctc ttcctagcta ctttcatcaa ttgtggaaga
      661 ttcagcgata ttaagaacgt tgatccgaaa tcatttaaat tagtccaaaa taagtatctg
      721 ggagtaataa tccagtgttt agtgacagag acaaagacaa gcgttagtag gcacatatac
      781 ttctttagcg caaggggtag gatcgatcca cttgtatatt tggatgaatt tttgaggaat
      841 tctgaaccag tcctaaaacg agtaaatagg accggcaatt cttcaagcaa taaacaggaa
      901 taccaattat taaaagataa cttagtcaga tcgtacaata aagctttgaa gaaaaatgcg
      961 ccttattcaa tctttgctat aaaaaatggc ccaaaatctc acattggaag acatttgatg
     1021 acctcatttc tttcaatgaa gggcctaacg gagttgacta atgttgtggg aaattggagc
     1081 gataagcgtg cttctgccgt ggccaggaca acgtatactc atcagataac agcaatacct
     1141 gatcactact tcgcactagt ttctcggtac tatgcatatg atccaatatc aaaggaaatg
     1201 atagcattga aggatgagac taatccaatt gaggagtggc agcatataga acagctaaag
     1261 ggtagtgctg aaggaagcat acgatacccc gcatggaatg ggataatatc acaggaggta
     1321 ctagactacc tttcatccta cataaataga cgcatataac catggcatgc atggtatcga
     1381 gatggcacat agccttgctc aaattggaat caggtttgtg ccaataccag tagaaacaga
     1441 cgaagaatcc atgggtatgg acagttttcc ctttgatatg taacggtgaa cagttgttct
     1501 acttttgttt gttagtcttg atgcttcact gatagataca agagccataa gaacctcaga
     1561 tccttccgta tttagccagt atgttctcta gtgtggttcg ttgtttttgc gtgagccatg
     1621 agaacgaacc attgagatca tacttacttt gcatgtcact caaaaatttt gcctcaaaac
     1681 tggtgagctg aatttttgca gttaaagcat cgtgtagtgt ttttcttagt ccgttacgta
     1741 ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc attcattttt atctggttgt
     1801 tctcaagttc ggttacgaga tccatttgtc tatctagttc aacttggaaa atcaacgtat
     1861 cagtcgggcg gcctcgctta tcaaccacca atttcatatt gctgtaagtg tttaaatctt
     1921 tacttattgg tttcaaaacc cattggttaa gccttttaaa ctcatggtag ttattttcaa
     1981 gcattaacat gaacttaaat tcatcaaggc taatctctat atttgccttg tgagttttct
     2041 tttgtgttag ttcttttaat aaccactcat aaatcctcat agagtatttg ttttcaaaag
     2101 acttaacatg ttccagatta tattttatga atttttttaa ctggaaaaga taaggcaata
     2161 tctcttcact aaaaactaat tctaattttt cgcttgagaa cttggcatag tttgtccact
     2221 ggaaaatctc aaagccttta accaaaggat tcctgatttc cacagttctc gtcatcagct
     2281 ctctggttgc tttagctaat acaccataag cattttccct actgatgttc atcatctgag
     2341 cgtattggtt ataagtgaac gataccgtcc gttctttcct tgtagggttt tcaatcgtgg
     2401 ggttgagtag tgccacacag cataaaatta gcttggtttc atgctccgtt aagtcatagc
     2461 gactaatcgc tagttcattt gctttgaaaa caactaattc agacatacat ctcaattggt
     2521 ctaggtgatt ttaatcacta taccaattga gatgggctag tcaatgataa ttactagtcc
     2581 ttttcctttg agttgtgggt atctgtaaat tctgctagac ctttgctgga aaacttgtaa
     2641 attctgctag accctctgta aattccgcta gacctttgtg tgtttttttt gtttatattc
     2701 aagtggttat aatttataga ataaagaaag aataaaaaaa gataaaaaga atagatccca
     2761 gccctgtgta taactcacta ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa
     2821 cgctgtttgc tcctctacaa aacagacctt aaaaccctaa aggcttaagt agcaccctcg
     2881 caagctcggt tgcggccgca atcgggcaaa tcgctgaata ttccttttgt ctccgaccat
     2941 caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca cggctctggc
     3001 agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa ggaaactacc
     3061 cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg gtctgctatg
     3121 tggtgctatc tgactttttg ctgttcagca gttcctgccc tctgattttc cagtctgacc
     3181 acttcggatt atcccgtgac aggtcattca gactggctaa tgcacccagt aaggcagcgg
     3241 tatcatcaac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3301 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3361 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3421 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3481 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3541 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3601 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3661 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3721 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3781 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3841 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3901 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3961 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4021 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4081 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4141 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4201 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4261 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4321 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4381 tgccacctg
//
