LOCUS       40924_17586        7518 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pEPIFOS-5, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7518)
  AUTHORS   EPICENTRE Biotechnologies.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-AUG-2007) 726 Post Road, Madison, WI 53713, USA
REFERENCE   2  (bases 1 to 7518)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7518)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (23-AUG-2007) 726 Post Road, Madison, WI 53713, USA"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     288..304
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        311..329
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(443..459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(467..483)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(491..521)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(536..557)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(780..1436)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1437..1539)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      2466..2685
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             2776..3528
                     /codon_start=1
                     /label=repE
                     /note="replication initiation protein for the bacterial F 
                     plasmid"
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    3534..3784
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             4110..5282
                     /codon_start=1
                     /label=sopA
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             5285..6253
                     /codon_start=1
                     /label=sopB
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    6329..6802
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     misc_feature    complement(7062..7460)
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     protein_bind    complement(7478..7511)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gcggccgcaa ggggttcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
       61 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
      121 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt cagctgcgca actgttggga
      181 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      241 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      301 cagtgaattg taatacgact cactataggg cgaattcgag ctcggtaccc ggggatccca
      361 cgtgggatcc tctagagtcg acctgcaggc atgcaagctt gagtattcta tagtctcacc
      421 taaatagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
      481 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
      541 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
      601 cgtgccagct gcattaatga atcggccaac gcgaacccct tgcggccgcc cgggccgtcg
      661 accaattctc atgtttgaca gcttatcatc gaatttctgc cattcatccg cttattatca
      721 cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta aaaaaattac
      781 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
      841 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
      901 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg
      961 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca
     1021 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat
     1081 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca
     1141 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
     1201 tctttcattg ccatacgaaa ttccggatga gcattcatca ggcgggcaag aatgtgaata
     1261 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc
     1321 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct
     1381 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta
     1441 gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt
     1501 tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccaaaag
     1561 ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc tgcgaagtga
     1621 tcttccgtca caggtattta ttcgcgataa gctcatggag cggcgtaacc gtcgcacagg
     1681 aaggacagag aaagcgcgga tctgggaagt gacggacaga acggtcagga cctggattgg
     1741 ggaggcggtt gccgccgctg ctgctgacgg tgtgacgttc tctgttccgg tcacaccaca
     1801 tacgttccgc cattcctatg cgatgcacat gctgtatgcc ggtataccgc tgaaagttct
     1861 gcaaagcctg atgggacata agtccatcag ttcaacggaa gtctacacga aggtttttgc
     1921 gctggatgtg gctgcccggc accgggtgca gtttgcgatg ccggagtctg atgcggttgc
     1981 gatgctgaaa caattatcct gagaataaat gccttggcct ttatatggaa atgtggaact
     2041 gagtggatat gctgtttttg tctgttaaac agagaagctg gctgttatcc actgagaagc
     2101 gaacgaaaca gtcgggaaaa tctcccatta tcgtagagat ccgcattatt aatctcagga
     2161 gcctgtgtag cgtttatagg aagtagtgtt ctgtcatgat gcctgcaagc ggtaacgaaa
     2221 acgatttgaa tatgccttca ggaacaatag aaatcttcgt gcggtgttac gttgaagtgg
     2281 agcggattat gtcagcaatg gacagaacaa cctaatgaac acagaaccat gatgtggtct
     2341 gtccttttac agccagtagt gctcgccgca gtcgagcgac agggcgaagc cctcgagtga
     2401 gcgaggaagc accagggaac agcacttata tattctgctt acacacgatg cctgaaaaaa
     2461 cttcccttgg ggttatccac ttatccacgg ggatattttt ataattattt tttttatagt
     2521 ttttagatct tcttttttag agcgccttgt aggcctttat ccatgctggt tctagagaag
     2581 gtgttgtgac aaattgccct ttcagtgtga caaatcaccc tcaaatgaca gtcctgtctg
     2641 tgacaaattg cccttaaccc tgtgacaaat tgccctcaga agaagctgtt ttttcacaaa
     2701 gttatccctg cttattgact cttttttatt tagtgtgaca atctaaaaac ttgtcacact
     2761 tcacatggat ctgtcatggc ggaaacagcg gttatcaatc acaagaaacg taaaaatagc
     2821 ccgcgaatcg tccagtcaaa cgacctcact gaggcggcat atagtctctc ccgggatcaa
     2881 aaacgtatgc tgtatctgtt cgttgaccag atcagaaaat ctgatggcac cctacaggaa
     2941 catgacggta tctgcgagat ccatgttgct aaatatgctg aaatattcgg attgacctct
     3001 gcggaagcca gtaaggatat acggcaggca ttgaagagtt tcgcggggaa ggaagtggtt
     3061 ttttatcgcc ctgaagagga tgccggcgat gaaaaaggct atgaatcttt tccttggttt
     3121 atcaaacgtg cgcacagtcc atccagaggg ctttacagtg tacatatcaa cccatatctc
     3181 attcccttct ttatcgggtt acagaaccgg tttacgcagt ttcggcttag tgaaacaaaa
     3241 gaaatcacca atccgtatgc catgcgttta tacgaatccc tgtgtcagta tcgtaagccg
     3301 gatggctcag gcatcgtctc tctgaaaatc gactggatca tagagcgtta ccagctgcct
     3361 caaagttacc agcgtatgcc tgacttccgc cgccgcttcc tgcaggtctg tgttaatgag
     3421 atcaacagca gaactccaat gcgcctctca tacattgaga aaaagaaagg ccgccagacg
     3481 actcatatcg tattttcctt ccgcgatatc acttccatga cgacaggata gtctgagggt
     3541 tatctgtcac agatttgagg gtggttcgtc acatttgttc tgacctactg agggtaattt
     3601 gtcacagttt tgctgtttcc ttcagcctgc atggattttc tcatactttt tgaactgtaa
     3661 tttttaagga agccaaattt gagggcagtt tgtcacagtt gatttccttc tctttccctt
     3721 cgtcatgtga cctgatatcg ggggttagtt cgtcatcatt gatgagggtt gattatcaca
     3781 gtttattact ctgaattggc tatccgcgtg tgtacctcta cctggagttt ttcccacggt
     3841 ggatatttct tcttgcgctg agcgtaagag ctatctgaca gaacagttct tctttgcttc
     3901 ctcgccagtt cgctcgctat gctcggttac acggctgcgg cgagcgctag tgataataag
     3961 tgactgaggt atgtgctctt cttatctcct tttgtagtgt tgctcttatt ttaaacaact
     4021 ttgcggtttt ttgatgactt tgcgattttg ttgttgcttt gcagtaaatt gcaagattta
     4081 ataaaaaaac gcaaagcaat gattaaagga tgttcagaat gaaactcatg gaaacactta
     4141 accagtgcat aaacgctggt catgaaatga cgaaggctat cgccattgca cagtttaatg
     4201 atgacagccc ggaagcgagg aaaataaccc ggcgctggag aataggtgaa gcagcggatt
     4261 tagttggggt ttcttctcag gctatcagag atgccgagaa agcagggcga ctaccgcacc
     4321 cggatatgga aattcgagga cgggttgagc aacgtgttgg ttatacaatt gaacaaatta
     4381 atcatatgcg tgatgtgttt ggtacgcgat tgcgacgtgc tgaagacgta tttccaccgg
     4441 tgatcggggt tgctgcccat aaaggtggcg tttacaaaac ctcagtttct gttcatcttg
     4501 ctcaggatct ggctctgaag gggctacgtg ttttgctcgt ggaaggtaac gacccccagg
     4561 gaacagcctc aatgtatcac ggatgggtac cagatcttca tattcatgca gaagacactc
     4621 tcctgccttt ctatcttggg gaaaaggacg atgtcactta tgcaataaag cccacttgct
     4681 ggccggggct tgacattatt ccttcctgtc tggctctgca ccgtattgaa actgagttaa
     4741 tgggcaaatt tgatgaaggt aaactgccca ccgatccaca cctgatgctc cgactggcca
     4801 ttgaaactgt tgctcatgac tatgatgtca tagttattga cagcgcgcct aacctgggta
     4861 tcggcacgat taatgtcgta tgtgctgctg atgtgctgat tgttcccacg cctgctgagt
     4921 tgtttgacta cacctccgca ctgcagtttt tcgatatgct tcgtgatctg ctcaagaacg
     4981 ttgatcttaa agggttcgag cctgatgtac gtattttgct taccaaatac agcaatagta
     5041 atggctctca gtccccgtgg atggaggagc aaattcggga tgcctgggga agcatggttc
     5101 taaaaaatgt tgtacgtgaa acggatgaag ttggtaaagg tcagatccgg atgagaactg
     5161 tttttgaaca ggccattgat caacgctctt caactggtgc ctggagaaat gctctttcta
     5221 tttgggaacc tgtctgcaat gaaattttcg atcgtctgat taaaccacgc tgggagatta
     5281 gataatgaag cgtgcgcctg ttattccaaa acatacgctc aatactcaac cggttgaaga
     5341 tacttcgtta tcgacaccag ctgccccgat ggtggattcg ttaattgcgc gcgtaggagt
     5401 aatggctcgc ggtaatgcca ttactttgcc tgtatgtggt cgggatgtga agtttactct
     5461 tgaagtgctc cggggtgata gtgttgagaa gacctctcgg gtatggtcag gtaatgaacg
     5521 tgaccaggag ctgcttactg aggacgcact ggatgatctc atcccttctt ttctactgac
     5581 tggtcaacag acaccggcgt tcggtcgaag agtatctggt gtcatagaaa ttgccgatgg
     5641 gagtcgccgt cgtaaagctg ctgcacttac cgaaagtgat tatcgtgttc tggttggcga
     5701 gctggatgat gagcagatgg ctgcattatc cagattgggt aacgattatc gcccaacaag
     5761 tgcttatgaa cgtggtcagc gttatgcaag ccgattgcag aatgaatttg ctggaaatat
     5821 ttctgcgctg gctgatgcgg aaaatatttc acgtaagatt attacccgct gtatcaacac
     5881 cgccaaattg cctaaatcag ttgttgctct tttttctcac cccggtgaac tatctgcccg
     5941 gtcaggtgat gcacttcaaa aagcctttac agataaagag gaattactta agcagcaggc
     6001 atctaacctt catgagcaga aaaaagctgg ggtgatattt gaagctgaag aagttatcac
     6061 tcttttaact tctgtgctta aaacgtcatc tgcatcaaga actagtttaa gctcacgaca
     6121 tcagtttgct cctggagcga cagtattgta taagggcgat aaaatggtgc ttaacctgga
     6181 caggtctcgt gttccaactg agtgtataga gaaaattgag gccattctta aggaacttga
     6241 aaagccagca ccctgatgcg accacgtttt agtctacgtt tatctgtctt tacttaatgt
     6301 cctttgttac aggccagaaa gcataactgg cctgaatatt ctctctgggc ccactgttcc
     6361 acttgtatcg tcggtctgat aatcagactg ggaccacggt cccactcgta tcgtcggtct
     6421 gattattagt ctgggaccac ggtcccactc gtatcgtcgg tctgattatt agtctgggac
     6481 cacggtccca ctcgtatcgt cggtctgata atcagactgg gaccacggtc ccactcgtat
     6541 cgtcggtctg attattagtc tgggaccatg gtcccactcg tatcgtcggt ctgattatta
     6601 gtctgggacc acggtcccac tcgtatcgtc ggtctgatta ttagtctgga accacggtcc
     6661 cactcgtatc gtcggtctga ttattagtct gggaccacgg tcccactcgt atcgtcggtc
     6721 tgattattag tctgggacca cgatcccact cgtgttgtcg gtctgattat cggtctggga
     6781 ccacggtccc acttgtattg tcgatcagac tatcagcgtg agactacgat tccatcaatg
     6841 cctgtcaagg gcaagtattg acatgtcgtc gtaacctgta gaacggagta acctcggtgt
     6901 gcggttgtat gcctgctgtg gattgctgct gtgtcctgct tatccacaac attttgcgca
     6961 cggttatgtg gacaaaatac ctggttaccc aggccgtgcc ggcacgttaa ccgggctgca
     7021 tccgatgcaa gtgtgtcgct gtcgacgagc tcgcgagctc ggacatgagg ttgccccgta
     7081 ttcagtgtcg ctgatttgta ttgtctgaag ttgtttttac gttaagttga tgcagatcaa
     7141 ttaatacgat acctgcgtca taattgatta tttgacgtgg tttgatggcc tccacgcacg
     7201 ttgtgatatg tagatgataa tcattatcac tttacgggtc ctttccggtg atccgacagg
     7261 ttacggggcg gcgacctcgc gggttttcgc tatttatgaa aattttccgg tttaaggcgt
     7321 ttccgttctt cttcgtcata acttaatgtt tttatttaaa ataccctctg aaaagaaagg
     7381 aaacgacagg tgctgaaagc gagctttttg gcctctgtcg tttcctttct ctgtttttgt
     7441 ccgtggaatg aacaatggaa gtccgagctc atcgctaata acttcgtata gcatacatta
     7501 tacgaagtta tattcgat
//
