LOCUS       sequence-69810-e        3630 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 3630)
  TITLE     Series-13: Co-transformation vectors for Series-2 transfer vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3630)
  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            "Unpublished"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 StrepII TEV co-transformation cloning vect...
FEATURES             Location/Qualifiers
     source          1..3630
                     /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             1347..1370
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1377..1397
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(1522..1540)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1536..1583
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1754..2545)
                     /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(2546..2637)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2685..3423)
                     /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 gctagctgga gccacccgca gttcgaaaaa ggcgccgaaa
     1381 acctgtactt ccaatccaat attggaagtg gataacggat ccgcgatcgc ggcgcgccac
     1441 ctggtggccg gccggtacca cgcgtgcgcg ctgatccggc tgctaacaaa gcccgaaagg
     1501 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     1561 aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac
     1621 tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac
     1681 cctgccctga accgacgacc gggtcatcgt ggccggatct tgcggcccct cggcttgaac
     1741 gaattgttag acattatttg ccgactacct tggtgatctc gcctttcacg taatggacaa
     1801 attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct
     1861 gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg
     1921 cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg
     1981 taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg
     2041 tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg
     2101 ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa
     2161 tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa
     2221 attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg
     2281 tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt
     2341 cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat
     2401 caatatcact gtgcggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca
     2461 gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata
     2521 cttcggcgat caccgcttcc ctcatactct tcctttttca ttattattga agcatttatc
     2581 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2641 ctagctcact cggtcgctac gctccgggcg tgagactgcg gcgggcgctg cggacacata
     2701 caaagttacc cacagattcc gtggataagc aggggactaa catgtgaggc aaaacagcag
     2761 ggccgcgccg gtggcgtttt tccataggct ccgccctcct gccagagttc acataaacag
     2821 acgcttttcc ggtgcatctg tgggagccgt gaggctcaac catgaatctg acagtacggg
     2881 cgaaacccga caggacttaa agatccccac cgtttccggc gggtcgctcc ctcttgcgct
     2941 ctcctgttcc gaccctgccg tttaccggat acctgttccg cctttctccc ttacgggaag
     3001 tgtggcgctt tctcatagct cacacactgg tatctcggct cggtgtaggt cgttcgctcc
     3061 aagctgggct gtaagcaaga actccccgtt cagcccgact gctgcgcctt atccggtaac
     3121 tgttcacttg agtccaaccc ggaaaagcac ggtaaaacgc cactggcagc agccattggt
     3181 aactgggagt tcgcagagga tttgtttagc taaacacgcg gttgctcttg aagtgtgcgc
     3241 caaagtccgg ctacactgga aggacagatt tggttgctgt gctctgcgaa agccagttac
     3301 cacggttaag cagttcccca actgacttaa ccttcgatca aaccacctcc ccaggtggtt
     3361 ttttcgttta cagggcaaaa gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3421 atcttttcta ctgaaccgct cttgatttca gtgcaattta tctcttcaaa tgtagcacct
     3481 gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc
     3541 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     3601 gagtgagcta acttacatta attgcgttgc
//
