LOCUS       sequence-69809-e        3990 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 3990)
  TITLE     Series-13: Co-transformation vectors for Series-2 transfer vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3990)
  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            "Unpublished"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 Mocr TEV co-transformation cloning ve...
FEATURES             Location/Qualifiers
     source          1..3990
                     /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             1737..1757
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(1882..1900)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1896..1943
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2114..2905)
                     /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(2906..2997)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3045..3783)
                     /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 gcaacatgac atacaataat gttttcgacc acgcgtacga aatgcttaag gagaacatcc
     1441 gttatgatga catacgggat acagatgacc ttcacgacgc aattcatatg gccgccgata
     1501 atgccgtccc tcactattat gccgatattc ggtcagttat ggcatctgaa ggtattgatc
     1561 tggaatttga ggatagtggc ctcatgccag atacgaagga cgacatcaga attctgcaag
     1621 cgcggatcta cgaacaactt accatcgatt tatgggaaga tgcggaagat ctcctgaatg
     1681 aatacctgga agaagtggag gagtatgagg aagatgaaga ggggacgggg atcgaggaaa
     1741 acctgtactt ccaatccaat attggaagtg gataacggat ccgcgatcgc ggcgcgccac
     1801 ctggtggccg gccggtacca cgcgtgcgcg ctgatccggc tgctaacaaa gcccgaaagg
     1861 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     1921 aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac
     1981 tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac
     2041 cctgccctga accgacgacc gggtcatcgt ggccggatct tgcggcccct cggcttgaac
     2101 gaattgttag acattatttg ccgactacct tggtgatctc gcctttcacg taatggacaa
     2161 attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct
     2221 gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg
     2281 cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg
     2341 taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg
     2401 tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg
     2461 ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa
     2521 tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa
     2581 attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg
     2641 tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt
     2701 cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat
     2761 caatatcact gtgcggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca
     2821 gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata
     2881 cttcggcgat caccgcttcc ctcatactct tcctttttca ttattattga agcatttatc
     2941 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3001 ctagctcact cggtcgctac gctccgggcg tgagactgcg gcgggcgctg cggacacata
     3061 caaagttacc cacagattcc gtggataagc aggggactaa catgtgaggc aaaacagcag
     3121 ggccgcgccg gtggcgtttt tccataggct ccgccctcct gccagagttc acataaacag
     3181 acgcttttcc ggtgcatctg tgggagccgt gaggctcaac catgaatctg acagtacggg
     3241 cgaaacccga caggacttaa agatccccac cgtttccggc gggtcgctcc ctcttgcgct
     3301 ctcctgttcc gaccctgccg tttaccggat acctgttccg cctttctccc ttacgggaag
     3361 tgtggcgctt tctcatagct cacacactgg tatctcggct cggtgtaggt cgttcgctcc
     3421 aagctgggct gtaagcaaga actccccgtt cagcccgact gctgcgcctt atccggtaac
     3481 tgttcacttg agtccaaccc ggaaaagcac ggtaaaacgc cactggcagc agccattggt
     3541 aactgggagt tcgcagagga tttgtttagc taaacacgcg gttgctcttg aagtgtgcgc
     3601 caaagtccgg ctacactgga aggacagatt tggttgctgt gctctgcgaa agccagttac
     3661 cacggttaag cagttcccca actgacttaa ccttcgatca aaccacctcc ccaggtggtt
     3721 ttttcgttta cagggcaaaa gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3781 atcttttcta ctgaaccgct cttgatttca gtgcaattta tctcttcaaa tgtagcacct
     3841 gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc
     3901 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     3961 gagtgagcta acttacatta attgcgttgc
//
