LOCUS       62056_21835        4755 bp DNA     circular SYN 12-MAY-2021
DEFINITION  Cloning vector pUC-2XproD, complete genome.
ACCESSION   MT333853
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4755)
  AUTHORS   Velasco L.
  TITLE     An efficient dsRNA constitutive expression system for bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4755)
  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 4755)
  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            "Unpublished"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..4755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      258..416
                     /label=proD
                     /note="proD"
                     /regulatory_class="promoter"
     protein_bind    423..547
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        572..602
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     gene            656..1336
                     /gene="cat"
                     /label=cat
     CDS             656..1336
                     /codon_start=1
                     /transl_table=11
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="QUV72830.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNEYNSTAMSGRAGRKRV
                     DPAY"
     CDS             1655..1957
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(2001..2125)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2140..2156)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(2173..2331)
                     /label=proD
                     /note="proD"
                     /regulatory_class="promoter"
     primer_bind     complement(2359..2375)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2849..2953
                     /label=AmpR promoter
     CDS             2954..3811
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3985..4573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gtcgactcta gagcacagct aacaccacgt cgtccctatc tgctgcccta
      301 ggtctatgag tggttgctgg ataactttac gggcatgcat aaggctcgta taatatattc
      361 agggagacca caacggtttc cctctacaaa taattttgtt taacttttac tagagtgata
      421 tcacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat
      481 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt
      541 cactatggcg gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa
      601 tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga
      661 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
      721 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
      781 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
      841 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
      901 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
      961 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     1021 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     1081 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     1141 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     1201 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     1261 cagaatgctt aatgaataca acagtactgc gatgagtggc agggcggggc gtaaacgcgt
     1321 ggatccggct tactaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta
     1381 taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca
     1441 gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc
     1501 aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa
     1561 cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac
     1621 ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa
     1681 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
     1741 gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga
     1801 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     1861 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     1921 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
     1981 cagccagtct gcaggtcgac catagtgact ggatatgttg tgttttacag tattatgtag
     2041 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
     2101 gttcagcttt cttgtacaaa gtggtgatat caagcttatc gataccgtcg acctcgaggg
     2161 ggggcccggt acctctagta aaagttaaac aaaattattt gtagagggaa accgttgtgg
     2221 tctccctgaa tatattatac gagccttatg catgcccgta aagttatcca gcaaccactc
     2281 atagacctag ggcagcagat agggacgacg tggtgttagc tgtgctctag aggatccccg
     2341 ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     2401 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     2461 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     2521 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     2581 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     2641 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2701 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     2761 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     2821 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2881 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2941 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3001 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3061 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3121 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3181 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3241 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3301 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3361 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3421 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3481 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3541 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3601 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3661 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3721 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3781 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3841 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3901 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3961 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4021 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4081 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4141 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4201 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4261 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4321 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4381 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4441 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4501 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4561 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4621 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4681 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     4741 cgaggaagcg gaaga
//
