LOCUS       sequence-69812-e        3966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3966)
  TITLE     Series-13: Co-transformation vectors for Series-2 transfer vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3966)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pET His6 Thioredoxin TEV co-transformation clo...
FEATURES             Location/Qualifiers
     source          1..3966
                     /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)"
     CDS             1350..1367
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1377..1703
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             1713..1733
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(1858..1876)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1872..1919
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2090..2881)
                     /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(2882..2973)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3021..3759)
                     /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 tatacatatg aaatcttctc accatcacca tcaccatggt tcttctatga
     1381 gcgataaaat tattcacctg actgacgaca gttttgacac ggatgtactc aaagcggacg
     1441 gggcgatcct cgtcgatttc tgggcagagt ggtgcggtcc gtgcaaaatg atcgccccga
     1501 ttctggatga aatcgctgac gaatatcagg gcaaactgac cgttgcaaaa ctgaacatcg
     1561 atcaaaaccc tggcactgcg ccgaaatatg gcatccgtgg tatcccgact ctgctgctgt
     1621 tcaaaaacgg tgaagtggcg gcaaccaaag tgggtgcact gtctaaaggt cagttgaaag
     1681 agttcctcga cgctaacctg gccgggatcg aggaaaacct gtacttccaa tccaatattg
     1741 gaagtggata acggatccgc gatcgcggcg cgccacctgg tggccggccg gtaccacgcg
     1801 tgcgcgctga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     1861 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     1921 tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg
     1981 taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt
     2041 catcgtggcc ggatcttgcg gcccctcggc ttgaacgaat tgttagacat tatttgccga
     2101 ctaccttggt gatctcgcct ttcacgtaat ggacaaattc ttccaactga tctgcgcgcg
     2161 aggccaagcg atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct
     2221 gatactgggc cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt
     2281 tgccggttac tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc
     2341 cagcccagtc gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat
     2401 cctgttcagg aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat
     2461 gttctcttgc ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga
     2521 tacctgcaag aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat
     2581 aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct
     2641 cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg
     2701 tttcatcaag ccttacggtc accgtaacca gcaaatcaat atcactgtgc ggcttcaggc
     2761 cgccatccac tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct
     2821 cgatgacgcc aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca
     2881 tactcttcct ttttcattat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2941 acatatttga atgtatttag aaaaataaac aaatagctag ctcactcggt cgctacgctc
     3001 cgggcgtgag actgcggcgg gcgctgcgga cacatacaaa gttacccaca gattccgtgg
     3061 ataagcaggg gactaacatg tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca
     3121 taggctccgc cctcctgcca gagttcacat aaacagacgc ttttccggtg catctgtggg
     3181 agccgtgagg ctcaaccatg aatctgacag tacgggcgaa acccgacagg acttaaagat
     3241 ccccaccgtt tccggcgggt cgctccctct tgcgctctcc tgttccgacc ctgccgttta
     3301 ccggatacct gttccgcctt tctcccttac gggaagtgtg gcgctttctc atagctcaca
     3361 cactggtatc tcggctcggt gtaggtcgtt cgctccaagc tgggctgtaa gcaagaactc
     3421 cccgttcagc ccgactgctg cgccttatcc ggtaactgtt cacttgagtc caacccggaa
     3481 aagcacggta aaacgccact ggcagcagcc attggtaact gggagttcgc agaggatttg
     3541 tttagctaaa cacgcggttg ctcttgaagt gtgcgccaaa gtccggctac actggaagga
     3601 cagatttggt tgctgtgctc tgcgaaagcc agttaccacg gttaagcagt tccccaactg
     3661 acttaacctt cgatcaaacc acctccccag gtggtttttt cgtttacagg gcaaaagatt
     3721 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctactga accgctcttg
     3781 atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag
     3841 ttgtaattct catgttagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     3901 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     3961 cgttgc
//
