LOCUS       62056_19510        7457 bp DNA     circular SYN 28-MAR-2020
DEFINITION  Cloning vector pSEVA2514-rec2, complete sequence.
ACCESSION   MN180223
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7457)
  AUTHORS   Aparicio T, Martinez-Garcia E, de Lorenzo V.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUL-2019) Systems Biology Program, Centro Nacional de 
            Biotecnologia, CSIC, Darwin 3, Madrid, Madrid 28049, Spain
REFERENCE   2  (bases 1 to 7457)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (15-JUL-2019) Systems Biology Program, Centro Nacional de 
            Biotecnologia, CSIC, Darwin 3, Madrid, Madrid 28049, Spain"
FEATURES             Location/Qualifiers
     source          1..7457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(12..722)
                     /label=lambda repressor (ts)
                     /note="temperature-sensitive variant of the phage lambda
                     repressor"
     regulatory      complement(729..752)
                     /label=tir region
                     /note="tir region"
                     /regulatory_class="other"
     regulatory      complement(753..858)
                     /label=PKan (Km promoter)
                     /note="PKan (Km promoter)"
                     /regulatory_class="promoter"
     primer_bind     1009..1032
                     /label=R24
                     /note="R24"
     regulatory      1033..1414
                     /label=PL promoter
                     /note="PL promoter"
                     /regulatory_class="promoter"
     gene            1455..2258
                     /gene="rec2"
                     /label=rec2
     CDS             1455..2258
                     /codon_start=1
                     /transl_table=11
                     /gene="rec2"
                     /product="Rec2"
                     /label=rec2
                     /note="Rec2 recombinase"
                     /protein_id="QIP75699.1"
                     /translation="MSNAALAERQNETQLLDAPAGAVQATESSMMIQMIQRAAADPAVD
                     VDKMERLMQMHERFVDRQASAAFNAAMVRAQRRIKPVARRALNVQTNSTYARLEDIDRE
                     ISPIFTEEGFSLSFGTGDSHLAGYIRVICDVMHDQGHTRQYKMDLPLDATGIGGKTNKT
                     GVHAHGSTNSYARRYLTMNIFNVVMANEDTDGNAEPPEEPVITSRQAAQLEALLKKCSP
                     TMAGLFIEKYGCASNVYKSEFDEVLAKLTRSANRTQQEPVDANHH"
     primer_bind     complement(2303..2326)
                     /label=F24
                     /note="F24"
     terminator      2337..2431
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2563..3375
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     oriT            3545..3653
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(3667..4515)
                     /label=RSF1010 RepC
                     /note="replication protein C of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             complement(4505..5341)
                     /label=RSF1010 RepA
                     /note="replication protein A of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             complement(5855..6823)
                     /label=RSF1010 RepB
                     /note="replication protein B of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     promoter        complement(6847..6877)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     rep_origin      6933..7327
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     terminator      complement(7365..7451)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaatc agccaaacgt ctcttcaggc cactgactag cgataacttt ccccacaacg
       61 gaacaactct cattgcatgg gatcattggg tactgtgggt ttagtggttg taaaaacacc
      121 tgaccgctat ccctgatcag tttcttgaag gtaaactcat cacccccaag tctggctatg
      181 cagaaatcac ctggctcaac agcctgctca gggtcaacga gaattaacat tccgtcagga
      241 aagctcggct tggagcctgt tggtgcggtc atggaattac cttcaacctc aagccagaat
      301 gcagaatcac tggctttttt ggttgtgctt acccatctct ccgcatcacc tttggtaaag
      361 gttctgagct taggtgagaa catccctgcc tgaacatgag aaaaaacagg gtactcatac
      421 tcacttctaa gtgacggctg catactaacc gcttcataca tctcgtagat ttctctggcg
      481 attgaagggc taaattcttc aacgctaact ttgagaattt ttgtaagcaa tgcggcgtta
      541 taagcattta atgcattgat gccattaaat aaagcaccaa cgcctgactg ccccatcccc
      601 atcttgtctg cgacagattc ctgggataag ccaagttcat ttttcttttt ttcataaatt
      661 gctttaaggc ggcgtgcgtc ctcaagctgc tcttgtgtta atggtttctt ttttgtgctc
      721 atgctagctt tttcctcctt ataaagttaa tcaacacccc ttgtattact gtttatgtaa
      781 gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga
      841 ttttgagaca caagacgtgg caaaaaacat tatccagaac gggagtgcgc cttgagcgac
      901 acgaattatg cagtgattta cgacctgcac agccatacca cagcttccga tggctgcctg
      961 acgccagaag cattggtgca ccgtgcagtc gatgataagc tgtcaaacag cggataacaa
     1021 tttcacacag gataacccgg cctcagcgcc gggttttctt tgcctcacga tcgcccccaa
     1081 aacacataac caattgtatt tattgaaaaa taaatagata caactcacta aacatagcaa
     1141 ttcagatctc tcacctacca aacaatgccc ccctgcaaaa aataaattca tataaaaaac
     1201 atacagataa ccatctgcgg tgataaatta tctctggcgg tgttgacata aataccactg
     1261 gcggtgatac tgagcacatc agcaggacgc actgaccacc atgaaggtga cgctcttaaa
     1321 aattaagccc tgaagaaggg cagcattcaa agcagaaggc tttggggtgt gtgatacgaa
     1381 acgaagcatt ggccgtaagt gcgattccgg attacctagg ccgcggccgc gcgaattcag
     1441 aaggagaata taccatgagc aatgcagcct tggccgaacg gcagaatgaa acccagcttc
     1501 tggacgctcc cgccggagcg gtacaagcca ccgaatcgtc catgatgatt caaatgattc
     1561 aacgcgccgc ggctgacccg gcagtggacg tcgacaagat ggagcgcctg atgcagatgc
     1621 acgagcgctt tgtcgaccgc caagcctcgg ctgctttcaa cgctgccatg gtccgcgcac
     1681 aacggcggat aaaaccggtt gcccgacggg cgcttaatgt tcagaccaat agcacctacg
     1741 cccgactgga agacatcgac cgagagatca gcccgatctt cactgaagaa ggtttctcgc
     1801 tttcgttcgg taccggcgac tcccatctgg ctggttacat tcgggtgatt tgcgacgtga
     1861 tgcatgacca ggggcacacc aggcaataca agatggacct gccgctggac gccaccggca
     1921 ttgggggtaa gacgaacaag accggcgtcc acgctcatgg atccaccaac agttacgccc
     1981 gccgctatct cacgatgaac atcttcaatg tcgtcatggc gaacgaggac accgacggca
     2041 acgcggagcc gcccgaagag cccgtcatca cctcccgcca ggcagcacag ctcgaggcgc
     2101 tgttgaaaaa atgcagccca acgatggctg gcctgttcat tgaaaagtat ggctgcgcgt
     2161 cgaacgtcta caaatccgag ttcgacgagg tactggcgaa gctcaccagg tcagccaatc
     2221 gcactcagca ggagcccgtt gatgcaaatc atcactgagg atcctctaga gtcgacctgc
     2281 aggcatgcaa gcttgcggcc gcgtcgtgac tgggaaaacc ctggcgacta gtcttggact
     2341 cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg
     2401 ttgccgccgg gcgtttttta ttggtgagaa tccaggggtc cccaataatt acgatttaaa
     2461 tttgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     2521 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcagcgt
     2581 gaaacgagct gtagccgtcc gcgtctgaac agcaacatgg atgcggatct gtatggctat
     2641 aaatgggcgc gtgataacgt gggtcagagc ggcgcgacca tttatcgtct gtatggcaaa
     2701 ccggatgcgc cggaactgtt tctgaaacat ggcaaaggca gcgtggcgaa cgatgtgacc
     2761 gatgaaatgg tgcgtctgaa ctggctgacc gaatttatgc cgctgccgac cattaaacat
     2821 tttattcgca ccccggatga tgcgtggctg ctgaccaccg cgattccggg caaaaccgcg
     2881 tttcaggtgc tggaagaata tccggatagc ggcgaaaaca ttgtggatgc gctggccgtg
     2941 tttctgcgtc gtctgcatag cattccggtg tgcaactgcc cgtttaacag cgatcgtgtg
     3001 tttcgtctgg cccaggcgca gagccgtatg aacaacggcc tggtggatgc gagcgatttt
     3061 gatgatgaac gtaacggctg gccggtggaa caggtgtgga aagaaatgca taaactgctg
     3121 ccgtttagcc cggatagcgt ggtgacccac ggcgatttta gcctggataa cctgattttc
     3181 gatgaaggca aactgattgg ctgcattgat gtgggccgtg tgggcattgc ggatcgttat
     3241 caggatctgg ccattctgtg gaactgcctg ggcgaattta gcccgagcct gcaaaaacgt
     3301 ctgtttcaga aatatggcat tgataatccg gatatgaaca aactgcaatt tcatctgatg
     3361 ctggatgaat ttttctaata attaattgga ccgcggtccg cgcgttgtcc ttttccgctg
     3421 cataaccctg cttcggggtc attatagcga ttttttcggt atatccatcc tttttcgcac
     3481 gatatacagg attttgccaa agggttcgtg tagactttcc ttggtgtatc caacggcgtc
     3541 agccgggcag gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc
     3601 cttattcgca cctggcggtg ctcaacggga atcctgctct gcgaggctgg ccgtaggccg
     3661 gcctcagcct gccgccttgg gccgggtgat gtcgtacttg cccgccgcga actcggttac
     3721 cgtccagccc agcgcgacca gctccggcaa cgcctcgcgc acccgctggc ggcgcttgcg
     3781 catggtcgaa ccactggcct ctgacggcca gacatagccg cacaaggtat ctatggaagc
     3841 cttgccggtt ttgccggggt cgatccagcc acacagccgc tggtgcagca ggcgggcggt
     3901 ttcgctgtcc agcgcccgca cctcgtccat gctgatgcgc acatgctggc cgccacccat
     3961 gacggcctgc gcgatcaagg ggttcagggc cacgtacagg cgcccgtccg cctcgtcgct
     4021 ggcgtactcc gacagcagcc gaaacccctg ccgcttgcgg ccattctggg cgatgatgga
     4081 taccttccaa aggcgctcga tgcagtcctg tatgtgcttg agcgccccac cactatcgac
     4141 ctctgccccg atttcctttg ccagcgcccg atagctacct ttgaccacat ggcattcagc
     4201 ggtgacggcc tcccacttgg gttccaggaa cagccggagc tgccgtccgc cttcggtctt
     4261 gggttccggg ccaagcacta ggccattagg cccagccatg gccaccagcc cttgcaggat
     4321 gcgcagatca tcagcgccca gcggctccgg gccgctgaac tcgatccgct tgccgtcgcc
     4381 gtagtcatac gtcacgtcca gcttgctgcg cttgcgctcg ccccgcttga gggcacggaa
     4441 caggccgggg gccagacagt gcgccgggtc gtgccggacg tggctgaggc tgtgcttgtt
     4501 cttaggcttc accacggggc acccccttgc tcttgcgctg cctctccagc acggcgggct
     4561 tgagcacccc gccgtcatgc cgcctgaacc accgatcagc gaacggtgcg ccatagttgg
     4621 ccttgctcac accgaagcgg acgaagaacc ggcgctggtc gtcgtccaca ccccattcct
     4681 cggcctcggc gctggtcatg ctcgacaggt aggactgcca gcggatgtta tcgaccagta
     4741 ccgagctgcc ccggctggcc tgctgctggt cgcctgcgcc catcatggcc gcgcccttgc
     4801 tggcatggtg caggaacacg atagagcacc cggtatcggc ggcgatggcc tccatgcgac
     4861 cgatgacctg ggccatgggg ccgctggcgt tttcttcctc gatgtggaac cggcgcagcg
     4921 tgtccagcac catcaggcgg cggccctcgg cggcgcgctt gaggccgtcg aaccactccg
     4981 gggccatgat gttgggcagg ctgccgatca gcggctggat cagcaggccg tcagccacgg
     5041 cttgccgttc ctcggcgctg aggtgcgccc caagggcgtg caggcggtga tgaatggcgg
     5101 tgggcgggtc ttcggcgggc aggtagatca ccgggccggt gggcagttcg cccacctcca
     5161 gcagatccgg cccgcctgca atctgtgcgg ccagttgcag ggccagcatg gatttaccgg
     5221 caccaccggg cgacaccagc gccccgaccg taccggccac catgttgggc aaaacgtagt
     5281 ccagcggtgg cggcgctgct gcgaacgcct ccagaatatt gataggctta tgggtagcca
     5341 ttgattgcct cctttgcagg cagttggtgg ttaggcgctg gcggggtcac tacccccgcc
     5401 ctgcgccgct ctgagttctt ccaggcactc gcgcagcgcc tcgtattcgt cgtcggtcag
     5461 ccagaacttg cgctgacgca tccctttggc cttcatgcgc tcggcatatc gcgcttggcg
     5521 tacagcgtca gggctggcca gcaggtcgcc ggtctgcttg tccttttggt ctttcatatc
     5581 agtcaccgag aaacttgccg gggccgaaag gcttgtcttc gcggaacaag gacaaggtgc
     5641 agccgtcaag gttaaggctg gccatatcag cgactgaaaa gcggccagcc tcggccttgt
     5701 ttgacgtata accaaagcca ccgggcaacc aatagccctt gtcacttttg atcaggtaga
     5761 ccgaccctga agcgcttttt tcgtattcca taaaaccccc ttctgtgcgt gagtactcat
     5821 agtataacag gcgtgagtac caacgcaagc actacatgct gaaatctggc ccgcccctgt
     5881 ccatgcctcg ctggcggggt gccggtgccc gtgccagctc ggcccgcgca agctggacgc
     5941 tgggcagacc catgaccttg ctgacggtgc gctcgatgta atccgcttcg tggccgggct
     6001 tgcgctctgc cagcgctggg ctggcctcgg ccatggcctt gccgatttcc tcggcactgc
     6061 ggccccggct ggccagcttc tgcgcggcga taaagtcgca cttgctgagg tcatcaccga
     6121 agcgcttgac cagcccggcc atctcgctgc ggtactcgtc cagcgccgtg cgccggtggc
     6181 ggctaagctg ccgctcgggc agttcgaggc tggccagcct gcgggccttc tcctgctgcc
     6241 gctgggcctg ctcgatctgc tggccagcct gctgcaccag cgccgggcca gcggtggcgg
     6301 tcttgccctt ggattcacgc agcagcaccc acggctgata accggcgcgg gtggtgtgct
     6361 tgtccttgcg gttggtgaag cccgccaagc ggccatagtg gcggctgtcg gcgctggccg
     6421 ggtcggcgtc gtactcgctg gccagcgtcc gggcaatctg cccccgaagt tcaccgcctg
     6481 cggcgtcggc caccttgacc catgcctgat agttcttcgg gctggtttcc actaccaggg
     6541 caggctcccg gccctcggct ttcatgtcat ccaggtcaaa ctcgctgagg tcgtccacca
     6601 gcaccagacc atgccgctcc tgctcggcgg gcctgatata cacgtcattg ccctgggcat
     6661 tcatccgctt gagccatggc gtgttctgga gcacttcggc ggctgaccat tcccggttca
     6721 tcatctggcc ggtggtggcg tccctgacgc cgatatcgaa gcgctcacag cccatggcct
     6781 tgagctgtcg gcctatggcc tgcaaagtcc tgtcgttctt catcgggcca ccaagcgatt
     6841 cccacacatt atacgagccg gaagcataaa gtgtaaagcc tagatccgaa ggatgagccg
     6901 ggctgaatga tcgaccgaga caggccctgc ggggctgcac acgcgccccc acccttcggg
     6961 tagggggaaa ggccgctaaa gcggctaaaa gcgctccagc gtatttctgc ggggtttggt
     7021 gtggggttta gcgggctttg cccgcctttc cccctgccgc gcagcggtgg ggcggtgtgt
     7081 agcctagcgc agcgaataga ccagctatcc ggcctctggc cgggcatatt gggcaagggc
     7141 agcagcgccc cacaagggcg ctgataaccg cgcctagtgg attattctta gataatcatg
     7201 gatggatttt tccaacaccc cgccagcccc cgcccctgct gggtttgcag gtttgggggc
     7261 gtgacagtta ttgcaggggt tcgtgacagt tattgcaggg gggcgtgaca gttattgcag
     7321 gggttcgtga cagttagggc gcgcccagct gtctagggcg gcggatttgt cctactcagg
     7381 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
     7441 cgttttattt gatgcct
//
