LOCUS       sequence-69806-e        3621 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 3621)
  TITLE     Series-13: Co-transformation vectors for Series-2 transfer vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3621)
  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            "Unpublished"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 TEV co-transformation cloning vector
            ...
FEATURES             Location/Qualifiers
     source          1..3621
                     /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             1368..1388
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(1513..1531)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1527..1574
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1745..2536)
                     /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(2537..2628)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2676..3414)
                     /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 tcaccatgaa aacctgtact
     1381 tccaatccaa tattggaagt ggataacgga tccgcgatcg cggcgcgcca cctggtggcc
     1441 ggccggtacc acgcgtgcgc gctgatccgg ctgctaacaa agcccgaaag gaagctgagt
     1501 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     1561 tgaggggttt tttgctgaaa cctcaggcat ttgagaagca cacggtcaca ctgcttccgg
     1621 tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga ccctgccctg
     1681 aaccgacgac cgggtcatcg tggccggatc ttgcggcccc tcggcttgaa cgaattgtta
     1741 gacattattt gccgactacc ttggtgatct cgcctttcac gtaatggaca aattcttcca
     1801 actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt
     1861 caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat
     1921 ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta
     1981 catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta
     2041 gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta
     2101 ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg
     2161 tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca aattgcagtt
     2221 cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta
     2281 cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca
     2341 aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac
     2401 tgtgcggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg
     2461 gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga
     2521 tcaccgcttc cctcatactc ttcctttttc attattattg aagcatttat cagggttatt
     2581 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata gctagctcac
     2641 tcggtcgcta cgctccgggc gtgagactgc ggcgggcgct gcggacacat acaaagttac
     2701 ccacagattc cgtggataag caggggacta acatgtgagg caaaacagca gggccgcgcc
     2761 ggtggcgttt ttccataggc tccgccctcc tgccagagtt cacataaaca gacgcttttc
     2821 cggtgcatct gtgggagccg tgaggctcaa ccatgaatct gacagtacgg gcgaaacccg
     2881 acaggactta aagatcccca ccgtttccgg cgggtcgctc cctcttgcgc tctcctgttc
     2941 cgaccctgcc gtttaccgga tacctgttcc gcctttctcc cttacgggaa gtgtggcgct
     3001 ttctcatagc tcacacactg gtatctcggc tcggtgtagg tcgttcgctc caagctgggc
     3061 tgtaagcaag aactccccgt tcagcccgac tgctgcgcct tatccggtaa ctgttcactt
     3121 gagtccaacc cggaaaagca cggtaaaacg ccactggcag cagccattgg taactgggag
     3181 ttcgcagagg atttgtttag ctaaacacgc ggttgctctt gaagtgtgcg ccaaagtccg
     3241 gctacactgg aaggacagat ttggttgctg tgctctgcga aagccagtta ccacggttaa
     3301 gcagttcccc aactgactta accttcgatc aaaccacctc cccaggtggt tttttcgttt
     3361 acagggcaaa agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3421 actgaaccgc tcttgatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc
     3481 cccatacgat ataagttgta attctcatgt tagtcatgcc ccgcgcccac cggaaggagc
     3541 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     3601 aacttacatt aattgcgttg c
//
