LOCUS       sequence-69800-e        3612 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3612)
  TITLE     Series-13: Co-transformation vectors for Series-2 transfer vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3612)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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            "Unpublished"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 StrepII TEV co-transformation cloning...
FEATURES             Location/Qualifiers
     source          1..3612
                     /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             1398..1421
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1428..1448
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(1573..1591)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1587..1634
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1864..2679)
                     /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(1864..2679)
                     /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     2587..2606
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2680..2771)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2789..3424)
                     /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(2934..2953)
                     /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 tgctagctgg agccacccgc agttcgaaaa aggcgccgaa aacctgtact
     1441 tccaatccaa tattggaagt ggataacgga tccgcgatcg cggcgcgcca cctggtggcc
     1501 ggccggtacc acgcgtgcgc gctgatccgg ctgctaacaa agcccgaaag gaagctgagt
     1561 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     1621 tgaggggttt tttgctgaaa cctcaggcat ttgagaagca cacggtcaca ctgcttccgg
     1681 tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga ccctgccctg
     1741 aaccgacgac aagctgacga ccgggtctcc gcaagtggca cttttcgggg aaatgtgcgc
     1801 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa
     1861 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     1921 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     1981 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     2041 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     2101 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     2161 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     2221 ttgcgcctga gcgagacgaa atacgcggtc gctgttaaaa ggacaattac aaacaggaat
     2281 cgaatgcaac cggcgcagga acactgccag cgcatcaact atattttcac ctgaatcagg
     2341 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     2401 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     2461 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     2521 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     2581 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     2641 cggcctagag caagacgttt cccgttgaat atggctcata ctcttccttt ttcattatta
     2701 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     2761 aaataaacaa ataggcatgc tagcgcagaa acgtcctaga agatgccagg aggatactta
     2821 gcagagagac aataaggccg gagcgaagcc gtttttccat aggctccgcc cccctgacga
     2881 acatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata
     2941 ccaggcgttt ccccctgatg gctccctctt gcgctctcct gttcccgtcc tgcggcgtcc
     3001 gtgttgtggt ggaggcttta cccaaatcac cacgtcccgt tccgtgtaga cagttcgctc
     3061 caagctgggc tgtgtgcaag aaccccccgt tcagcccgac tgctgcgcct tatccggtaa
     3121 ctatcatctt gagtccaacc cggaaagaca cgacaaaacg ccactggcag cagccattgg
     3181 taactgagaa ttagtggatt tagttatcga gagtcttgaa gtggtggcct aacagaggct
     3241 acactgaaag gacagtattt ggtatctgcg ctccactaaa gccagttacc aggttaagca
     3301 gttccccaac tgacttaacc ttcgatcaaa ccgcctcccc aggcggtttt ttcgtttaca
     3361 gagcaggaga ttacgacgat cgtaaaagga tctcaagaag atcctttacg gattcccgac
     3421 accatcactc ttgatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc
     3481 catacgatat aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg
     3541 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     3601 cttacattaa tt
//
