LOCUS       sequence-71718-e        3576 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 3576)
  TITLE     Series-13: Co-transformation vectors for Series-2 transfer vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3576)
  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            "Unpublished"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 StrepII TEV co-transformation cloning vect...
FEATURES             Location/Qualifiers
     source          1..3576
                     /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             1362..1385
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1392..1412
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(1537..1555)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1551..1598
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1828..2643)
                     /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(1828..2643)
                     /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     2551..2570
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2644..2735)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2753..3388)
                     /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(2898..2917)
                     /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 atatggctag ctggagccac ccgcagttcg
     1381 aaaaaggcgc cgaaaacctg tacttccaat ccaatattgg aagtggataa cggatccgcg
     1441 atcgcggcgc gccacctggt ggccggccgg taccacgcgt gcgcgctgat ccggctgcta
     1501 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     1561 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaacctcag gcatttgaga
     1621 agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag
     1681 cggctattta acgaccctgc cctgaaccga cgacaagctg acgaccgggt ctccgcaagt
     1741 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1801 aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg
     1861 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     1921 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     1981 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     2041 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat
     2101 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     2161 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc ggtcgctgtt
     2221 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     2281 aactatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     2341 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     2401 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     2461 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     2521 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     2581 agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct
     2641 catactcttc ctttttcatt attattgaag catttatcag ggttattgtc tcatgagcgg
     2701 atacatattt gaatgtattt agaaaaataa acaaataggc atgctagcgc agaaacgtcc
     2761 tagaagatgc caggaggata cttagcagag agacaataag gccggagcga agccgttttt
     2821 ccataggctc cgcccccctg acgaacatca cgaaatctga cgctcaaatc agtggtggcg
     2881 aaacccgaca ggactataaa gataccaggc gtttccccct gatggctccc tcttgcgctc
     2941 tcctgttccc gtcctgcggc gtccgtgttg tggtggaggc tttacccaaa tcaccacgtc
     3001 ccgttccgtg tagacagttc gctccaagct gggctgtgtg caagaacccc ccgttcagcc
     3061 cgactgctgc gccttatccg gtaactatca tcttgagtcc aacccggaaa gacacgacaa
     3121 aacgccactg gcagcagcca ttggtaactg agaattagtg gatttagtta tcgagagtct
     3181 tgaagtggtg gcctaacaga ggctacactg aaaggacagt atttggtatc tgcgctccac
     3241 taaagccagt taccaggtta agcagttccc caactgactt aaccttcgat caaaccgcct
     3301 ccccaggcgg ttttttcgtt tacagagcag gagattacga cgatcgtaaa aggatctcaa
     3361 gaagatcctt tacggattcc cgacaccatc actcttgatt tcagtgcaat ttatctcttc
     3421 aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg
     3481 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     3541 cccggtgcct aatgagtgag ctaacttaca ttaatt
//
