LOCUS       40924_39028        7408 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSCT32, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7408)
  AUTHORS   Chen S.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-OCT-2016) School of Life Sciences, Qingdao 
            Agricultural University, 700 Changcheng Road, Qingdao, Shandong 
            266109, PR China
REFERENCE   2  (bases 1 to 7408)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7408)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (30-OCT-2016) School of Life Sciences, Qingdao Agricultural 
            University, 700 Changcheng Road, Qingdao, Shandong 266109, PR China"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7408
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..103)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      420..1008
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(1256..1684)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1982..2059
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2060..3139
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    3411..3427
                     /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)."
     CDS             3453..4112
                     /codon_start=1
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
                     /translation="DVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTK
                     GGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQ
                     DSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMRLKLK
                     DGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHSTGA"
     CDS             4113..4130
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      4353..4439
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4531..4558
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(4703..5299)
                     /codon_start=1
                     /product="TraI"
                     /label=TraI
                     /protein_id="ARN18289.1"
                     /translation="MRSIKKSDFAELVKYITDEQGKTERLGHVRVTNCEANTLPAVMAE
                     VMATQHGNTRSEADKTYHLLVSFRAGEKPDAETLRAIEDRICAGLGFAEHQRVSAVHHD
                     TDNLHIHIAINKIHPTRNTIHEPYRAYRALADLCATLERDYGLERDNHETRQRVSENRA
                     NDMERHAGVESLVGWIIYSASQSLIKTVSASLIQM"
     CDS             complement(5358..5726)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(5759..5868)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(6752..7408)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
       61 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca
      121 aaagttggcc cagggcttcc cggtatcaac agggacacca ggatttattt attctgcgaa
      181 gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg tcgggtgatg ctgccaactt
      241 actgatttag tgtatgatgg tgtttttgag gtgctccagt ggcttctgtt tctatcagct
      301 gtccctcctg ttcagctact gacggggtgg tgcgtaacgg caaacatgac caaaatccct
      361 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
      421 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
      481 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
      541 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
      601 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
      661 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
      721 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      781 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      841 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      901 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      961 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     1021 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     1081 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     1141 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     1201 cggtattttc tccttacgca tctgtgcggt atttcacacc gcataatttt gttaaaattc
     1261 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     1321 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     1381 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     1441 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     1501 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     1561 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     1621 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     1681 gcgtcccatt cgccattcag gctgctatgg tgcactctca gtacaatctg ctctgatgcc
     1741 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     1801 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     1861 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     1921 cgaaacgcgc gaggcagcag atcaattcgc gcgcgaaggc gaagcggcat gcatttacgt
     1981 tgacaccatc gaatggtgca aaacctttcg cggtatggca tgatagcgcc cggaagagag
     2041 tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
     2101 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
     2161 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca
     2221 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca
     2281 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt
     2341 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct
     2401 tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat
     2461 tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac
     2521 acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct
     2581 ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc
     2641 gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc
     2701 ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa
     2761 tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat
     2821 gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga
     2881 cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg
     2941 cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa
     3001 gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cacccaatac
     3061 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     3121 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagcgc gaattgatct
     3181 ggtttgacag cttatcatcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat
     3241 cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg
     3301 cgcactcccg ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt
     3361 ctgaaatgag ctgttgacaa ttaatcatcc ggtccgtata atctgtggaa ttgtgagcgg
     3421 ataacaattt cacacaggaa acagaccata tggacgtcat caaggagttc atgcgcttca
     3481 aggtgcgcat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     3541 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc
     3601 ccttcgcctg ggacatcctg tcccctcagt tccagtacgg ctccaaggcc tacgtgaagc
     3661 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     3721 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     3781 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     3841 taatgcagaa gaagaccatg ggctgggagg cctccaccga gcggatgtac cccgaggacg
     3901 gcgccctgaa gggcgagatc aagatgaggc tgaagctgaa ggacggcggc cactacgacg
     3961 ccgaggtcaa gaccacctac atggccaaga agcccgtgca gctgcccggc gcctacaaga
     4021 ccgacatcaa gctggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     4081 agcgcgccga gggccgccac tccaccggcg cccatcatca tcaccatcac taagatctgg
     4141 taccaagctt ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc
     4201 agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc
     4261 acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc
     4321 tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag
     4381 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     4441 cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc
     4501 cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg
     4561 cgtttctaca aactcttttt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4621 gtacccacgc cgaaacaagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc
     4681 tgctggctac cctgtggaac acctacatct gtattaacga agcgctaacc gtttttatca
     4741 ggctctggga ggcagaataa atgatccagc cgaccaggct ttccacgccc gcgtgccgct
     4801 ccatgtcgtt cgcgcggttc tcggaaacgc gctgccgcgt ttcgtgattg tcacgctcaa
     4861 gcccgtagtc ccgttcgagc gtcgcgcaga ggtcagcgag ggcgcggtag gcccgatacg
     4921 gctcatggat ggtgtttcgg gtcgggtgaa tcttgttgat ggcgatatgg atgtgcaggt
     4981 tgtcggtgtc gtgatgcacg gcactgacgc gctgatgctc ggcgaagcca agcccagcgc
     5041 agatgcggtc ctcaatcgcg cgcaacgtct ccgcgtcggg cttctctccc gcgcggaagc
     5101 taaccagcag gtgataggtc ttgtcggcct cggaacgggt gttgccgtgc tgggtcgcca
     5161 tcacctcggc catgacagcg ggcagggtgt ttgcctcgca gttcgtgacg cgcacgtgac
     5221 ccaggcgctc ggtcttgcct tgctcgtcgg tgatgtactt caccagctcc gcgaagtcgc
     5281 tcttcttgat ggagcgcatg gggacgtgct tggcaatcac gcgcaccccc cggccgtttt
     5341 agcggctaaa aaagtcatgg ctctgccctc gggcggacca cgcccatcat gaccttgcca
     5401 agctcgtcct gcttctcttc gatcttcgcc agcagggcga ggatcgtggc atcaccgaac
     5461 cgcgccgtgc gcgggtcgtc ggtgagccag agtttcagca ggccgcccag gcggcccagg
     5521 tcgccattga tgcgggccag ctcgcggacg tgctcatagt ccacgacgcc cgtgattttg
     5581 tagccctggc cgacggccag caggtaggcc gacaggctca tgccggccgc cgccgccttt
     5641 tcctcaatcg ctcttcgttc gtctggaagg cagtacacct tgataggtgg gctgcccttc
     5701 ctggttggct tggtttcatc agccatccgc ttgccctcat ctgttacgcc ggcggtagcc
     5761 ggccagcctc gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg
     5821 aagaaggaac acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt
     5881 tggatacacc aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa
     5941 aaggatggat ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa
     6001 aagcgctgct tccctgctgt tttgtggaat atctaccgac tggaaacagg caaatgcagg
     6061 aaattactga actgagggga caggcgagag acgatgccaa agagctacac cgacgagctg
     6121 gccgagtggg ttgaatcccg cgcggccaag aagcgccggc gtgatgaggc tgcggttgcg
     6181 ttcctggcgg tgagggcgga tgtcgaggcg gcgttagcgt ccggctatgc gctcgtcacc
     6241 atttgggagc acatgcggga aacggggaag gtcaagttct cctacgagac gttccgctcg
     6301 cacgccaggc ggcacatcaa ggccaagccc gccgatgtgc ccgcaccgca ggccaaggct
     6361 gcggaacccg cgccggcacc caagacgccg gagccacggc ggccgaagca ggggggcaag
     6421 gctgaaaagc cggcccccgc tgcggccccg accggcttca ccttcaaccc aacaccggac
     6481 aaaaaggatc atatcgtcaa ttattacctc cacggggaga gcctgagcaa actggcctca
     6541 ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca
     6601 atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct
     6661 ttcgaatttc tgccattcat ccgcttatta tcacttattc aggcgtagca ccaggcgttt
     6721 aagggcacca ataactgcct taaaaaaatt acgccccgcc ctgccactca tcgcagtact
     6781 gttgtaattc attaagcatt ctgccgacat ggaagccatc acagacggca tgatgaacct
     6841 gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa
     6901 cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc
     6961 agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt
     7021 tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt
     7081 ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag
     7141 ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg aattccggat
     7201 gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt
     7261 tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt
     7321 gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg
     7381 tggtatatcc agtgattttt ttctccat
//
