LOCUS       pET co-transformation cloning vector (13S-A)        3566 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3566)
  TITLE     Series-13: Co-transformation vectors for Series-2 transfer vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3566)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            //gt12365.cn
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pET co-transformation cloning vector (13S-A)
FEATURES             Location/Qualifiers
     source          1..3566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(3..1085)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1047..1066
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1086..1163)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1209..1228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1209..1227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1232..1255
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     RBS             1308..1330
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1458..1476)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1472..1519
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1690..2481)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2482..2573)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2621..3359)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
ORIGIN
        1 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
       61 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
      121 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
      181 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
      241 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc
      301 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
      361 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
      421 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
      481 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
      541 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
      601 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
      661 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
      721 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
      781 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
      841 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
      901 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
      961 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     1021 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     1081 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     1141 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     1201 taggaaatta atacgactca ctatagggag agaattgtga gcggataaca attcccctgt
     1261 agattaatta agcggccgcc ctgcaggact cgagttctag aaataatttt gtttaacttt
     1321 aagaaggaga tatagatatc ccaactccat aaggatccgc gatcgcggcg cgccacctgg
     1381 tggccggccg gtaccacgcg tgcgcgctga tccggctgct aacaaagccc gaaaggaagc
     1441 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1501 ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct
     1561 tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg
     1621 ccctgaaccg acgaccgggt catcgtggcc ggatcttgcg gcccctcggc ttgaacgaat
     1681 tgttagacat tatttgccga ctaccttggt gatctcgcct ttcacgtaat ggacaaattc
     1741 ttccaactga tctgcgcgcg aggccaagcg atcttcttct tgtccaagat aagcctgtct
     1801 agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc agtcggcagc
     1861 gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg acaacgtaag
     1921 cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg ttaaggtttc
     1981 atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct ccgccgctgg
     2041 acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca gatcaatgtc
     2101 gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt ctccaaattg
     2161 cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac
     2221 ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt
     2281 gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca gcaaatcaat
     2341 atcactgtgc ggcttcaggc cgccatccac tgcggagccg tacaaatgta cggccagcaa
     2401 cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag tcgatacttc
     2461 ggcgatcacc gcttccctca tactcttcct ttttcattat tattgaagca tttatcaggg
     2521 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaatagctag
     2581 ctcactcggt cgctacgctc cgggcgtgag actgcggcgg gcgctgcgga cacatacaaa
     2641 gttacccaca gattccgtgg ataagcaggg gactaacatg tgaggcaaaa cagcagggcc
     2701 gcgccggtgg cgtttttcca taggctccgc cctcctgcca gagttcacat aaacagacgc
     2761 ttttccggtg catctgtggg agccgtgagg ctcaaccatg aatctgacag tacgggcgaa
     2821 acccgacagg acttaaagat ccccaccgtt tccggcgggt cgctccctct tgcgctctcc
     2881 tgttccgacc ctgccgttta ccggatacct gttccgcctt tctcccttac gggaagtgtg
     2941 gcgctttctc atagctcaca cactggtatc tcggctcggt gtaggtcgtt cgctccaagc
     3001 tgggctgtaa gcaagaactc cccgttcagc ccgactgctg cgccttatcc ggtaactgtt
     3061 cacttgagtc caacccggaa aagcacggta aaacgccact ggcagcagcc attggtaact
     3121 gggagttcgc agaggatttg tttagctaaa cacgcggttg ctcttgaagt gtgcgccaaa
     3181 gtccggctac actggaagga cagatttggt tgctgtgctc tgcgaaagcc agttaccacg
     3241 gttaagcagt tccccaactg acttaacctt cgatcaaacc acctccccag gtggtttttt
     3301 cgtttacagg gcaaaagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3361 tttctactga accgctcttg atttcagtgc aatttatctc ttcaaatgta gcacctgaag
     3421 tcagccccat acgatataag ttgtaattct catgttagtc atgccccgcg cccaccggaa
     3481 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     3541 gagctaactt acattaattg cgttgc
//