LOCUS       62056_1390        4705 bp DNA     circular SYN 12-MAY-2021
DEFINITION  Cloning vector L4440-2XproD-gtwy, complete genome.
ACCESSION   MT333852
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4705)
  AUTHORS   Velasco L.
  TITLE     An efficient dsRNA constitutive expression system for bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4705)
  AUTHORS   Velasco L.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-APR-2020) Plant Protection, Instituto Andaluz de 
            Investigacion y Formacion Agraria y Pesquera (IFAPA), Cortijo de la 
            Cruz s/n, Churriana, Malaga 29140, Spain
REFERENCE   3  (bases 1 to 4705)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-APR-2020) Plant Protection, Instituto Andaluz de Investigacion y
            Formacion Agraria y Pesquera (IFAPA), Cortijo de la Cruz s/n, 
            Churriana, Malaga 29140, Spain"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        587..605
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      616..775
                     /label=proD
                     /note="proD"
                     /regulatory_class="promoter"
     protein_bind    783..907
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        932..962
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     gene            1016..1696
                     /gene="cat"
                     /label=cat
     CDS             1016..1696
                     /codon_start=1
                     /transl_table=11
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="CAT marker"
                     /protein_id="QUV72827.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNEYNSTAMSGRAGRKRV
                     DPAY"
     CDS             2015..2317
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(2361..2485)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2500..2516)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2706..2724)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2734..2750)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2892..3347
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3373..3477
                     /label=AmpR promoter
     CDS             3478..4335
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4509..4705
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
       61 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
      121 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
      181 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
      241 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      301 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      361 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      421 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
      481 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      541 agcgcagcga gtcagtgagc gaggaagcaa cctggcttat cgaaattaat acgactcact
      601 atagggagac cggcattcta gagcacagct aacaccacgt cgtccctatc tgctgcccta
      661 ggtctatgag tggttgctgg ataactttac gggcatgcat aaggctcgta taatatattc
      721 agggagacca caacggtttc cctctacaaa taattttgtt taacttttac tagagtgata
      781 tcacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat
      841 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt
      901 cactatggcg gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa
      961 tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga
     1021 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     1081 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     1141 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     1201 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
     1261 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     1321 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     1381 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     1441 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     1501 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     1561 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     1621 cagaatgctt aatgaataca acagtactgc gatgagtggc agggcggggc gtaaacgcgt
     1681 ggatccggct tactaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta
     1741 taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca
     1801 gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc
     1861 aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa
     1921 cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac
     1981 ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa
     2041 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
     2101 gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga
     2161 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     2221 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     2281 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
     2341 cagccagtct gcaggtcgac catagtgact ggatatgttg tgttttacag tattatgtag
     2401 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
     2461 gttcagcttt cttgtacaaa gtggtgatat caagcttatc gataccgtcg acctcgaggg
     2521 ggggcccggt acctctagta aaagttaaac aaaattattt gtagagggaa accgttgtgg
     2581 tctccctgaa tatattatac gagccttatg catgcccgta aagttatcca gcaaccactc
     2641 atagacctag ggcagcagat agggacgacg tggtgttagc tgtgctctag aagtacccaa
     2701 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     2761 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2821 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2881 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2941 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3001 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3061 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3121 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3181 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3241 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3301 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     3361 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3421 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3481 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3541 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3601 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3661 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3721 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3781 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3841 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3901 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3961 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4021 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4081 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4141 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4201 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4261 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4321 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4381 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4441 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4501 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4561 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4621 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4681 caccacttca agaactctgt agcac
//
