LOCUS       sequence-69803-e        3885 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 3885)
  TITLE     Series-13: Co-transformation vectors for Series-2 transfer vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3885)
  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            "Unpublished"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 Sumo TEV co-transformation cloning ve...
FEATURES             Location/Qualifiers
     source          1..3885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(10..1092)
                     /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     1054..1073
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1093..1170)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1216..1235
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1216..1234
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1323..1345
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1365..1382
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1401..1694
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             1701..1721
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(1846..1864)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1860..1907
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2137..2952)
                     /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(2137..2952)
                     /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     2860..2879
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2953..3044)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3062..3697)
                     /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(3207..3226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
       61 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
      121 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
      181 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
      241 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc
      301 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
      361 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
      421 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
      481 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
      541 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
      601 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
      661 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
      721 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
      781 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
      841 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
      901 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
      961 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     1021 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     1081 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     1141 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
     1201 cctgcattag gaaattaata cgactcacta tagggagacc acaacggttt ccctctagtg
     1261 ccggctccgg agagctcttt aattaagcgg ccgccctgca ggactcgagt tctagaaata
     1321 attttgttta actttaagaa ggagatatac atatgaaatc ttctcaccat caccatcacc
     1381 atggttcttc tatggctagc atgtcggact cagaagtcaa tcaagaagct aagccagagg
     1441 tcaagccaga agtcaagcct gagactcaca tcaatttaaa ggtgtccgat ggatcttcag
     1501 agatcttctt caagatcaaa aagaccactc ctttaagaag gctgatggaa gcgttcgcta
     1561 aaagacaggg taaggaaatg gactccttaa gattcttgta cgacggtatt agaattcaag
     1621 ctgatcagac ccctgaagat ttggacatgg aggataacga tattattgag gctcacagag
     1681 aacagattgg tgggatcgag gaaaacctgt acttccaatc caatattgga agtggataac
     1741 ggatccgcga tcgcggcgcg ccacctggtg gccggccggt accacgcgtg cgcgctgatc
     1801 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     1861 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg
     1921 catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat
     1981 agacataagc ggctatttaa cgaccctgcc ctgaaccgac gacaagctga cgaccgggtc
     2041 tccgcaagtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2101 atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa
     2161 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     2221 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     2281 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     2341 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     2401 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     2461 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     2521 gtcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     2581 cagcgcatca actatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     2641 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     2701 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     2761 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     2821 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     2881 atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg
     2941 aatatggctc atactcttcc tttttcatta ttattgaagc atttatcagg gttattgtct
     3001 catgagcgga tacatatttg aatgtattta gaaaaataaa caaataggca tgctagcgca
     3061 gaaacgtcct agaagatgcc aggaggatac ttagcagaga gacaataagg ccggagcgaa
     3121 gccgtttttc cataggctcc gcccccctga cgaacatcac gaaatctgac gctcaaatca
     3181 gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg atggctccct
     3241 cttgcgctct cctgttcccg tcctgcggcg tccgtgttgt ggtggaggct ttacccaaat
     3301 caccacgtcc cgttccgtgt agacagttcg ctccaagctg ggctgtgtgc aagaaccccc
     3361 cgttcagccc gactgctgcg ccttatccgg taactatcat cttgagtcca acccggaaag
     3421 acacgacaaa acgccactgg cagcagccat tggtaactga gaattagtgg atttagttat
     3481 cgagagtctt gaagtggtgg cctaacagag gctacactga aaggacagta tttggtatct
     3541 gcgctccact aaagccagtt accaggttaa gcagttcccc aactgactta accttcgatc
     3601 aaaccgcctc cccaggcggt tttttcgttt acagagcagg agattacgac gatcgtaaaa
     3661 ggatctcaag aagatccttt acggattccc gacaccatca ctcttgattt cagtgcaatt
     3721 tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt aattctcatg
     3781 ttagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     3841 ggtcgagatc ccggtgccta atgagtgagc taacttacat taatt
//
