LOCUS       sequence-169629-        3567 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 3567)
  TITLE     Series-13: Co-transformation vectors for Series-2 transfer vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3567)
  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            "Unpublished"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..3567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     735..754
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(774..851)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     897..916
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        897..915
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1004..1026
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1046..1063
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1064..1084
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(1209..1227)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1223..1270
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1500..2315)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1500..2315)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2223..2242
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2316..2407)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2425..3060)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(2570..2589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3258..3567,1..773))
                     /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
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVDRSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDIEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
       61 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
      121 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
      181 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
      241 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
      301 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
      361 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
      421 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcggtctacc
      481 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
      541 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
      601 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
      661 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
      721 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
      781 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
      841 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaaattaat
      901 acgactcact atagggagac cacaacggtt tccctctagt gccggctccg gagagctctt
      961 taattaagcg gccgccctgc aggactcgag ttctagaaat aattttgttt aactttaaga
     1021 aggagatata catatgaaat cttctcacca tcaccatcac catgaaaacc tgtacttcca
     1081 atccaatatt ggaagtggat aacggatccg cgatcgcggc gcgccacctg gtggccggcc
     1141 ggtaccacgc gtgcgcgctg atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     1201 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     1261 gggttttttg ctgaaacctc aggcatttga gaagcacacg gtcacactgc ttccggtagt
     1321 caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc
     1381 gacgacaagc tgacgaccgg gtctccgcaa gtggcacttt tcggggaaat gtgcgcggaa
     1441 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct
     1501 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     1561 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     1621 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     1681 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     1741 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     1801 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     1861 gcctgagcga gacgaaatac gcggtcgctg ttaaaaggac aattacaaac aggaatcgaa
     1921 tgcaaccggc gcaggaacac tgccagcgca tcaactatat tttcacctga atcaggatat
     1981 tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca
     2041 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     2101 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     2161 aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca
     2221 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     2281 ctagagcaag acgtttcccg ttgaatatgg ctcatactct tcctttttca ttattattga
     2341 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     2401 aaacaaatag gcatgctagc gcagaaacgt cctagaagat gccaggagga tacttagcag
     2461 agagacaata aggccggagc gaagccgttt ttccataggc tccgcccccc tgacgaacat
     2521 cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag
     2581 gcgtttcccc ctgatggctc cctcttgcgc tctcctgttc ccgtcctgcg gcgtccgtgt
     2641 tgtggtggag gctttaccca aatcaccacg tcccgttccg tgtagacagt tcgctccaag
     2701 ctgggctgtg tgcaagaacc ccccgttcag cccgactgct gcgccttatc cggtaactat
     2761 catcttgagt ccaacccgga aagacacgac aaaacgccac tggcagcagc cattggtaac
     2821 tgagaattag tggatttagt tatcgagagt cttgaagtgg tggcctaaca gaggctacac
     2881 tgaaaggaca gtatttggta tctgcgctcc actaaagcca gttaccaggt taagcagttc
     2941 cccaactgac ttaaccttcg atcaaaccgc ctccccaggc ggttttttcg tttacagagc
     3001 aggagattac gacgatcgta aaaggatctc aagaagatcc tttacggatt cccgacacca
     3061 tcactcttga tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata
     3121 cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg
     3181 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     3241 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3301 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     3361 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     3421 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     3481 gttaacggcg ggatataaca tgagctgtct tcgatatcgt cgtatcccac taccgagatg
     3541 tccgcaccaa cgcgcagccc ggactcg
//
