pET28a-TmTrpB-2F3-opt

LOCUS       pET28a-TmTrpB-2F3-opt            6460 bp DNA     circular SYN 06-JUL-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    TmTrpB-Tm2F3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6460)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Sep 28, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      834..1422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1608..1750
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(1852..2043)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2852..3934)
                     /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"
     promoter        complement(3935..4012)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        4321..4339
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4340..4364
                     /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             4379..4401
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4420..4437
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4447..4464
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4468..5634
                     /codon_start=1
                     /label=Tm2F3
                     /translation="MKGYFGPYGGQYVPEILMGALEELEAAYEEIMKDESFWKEFNDLL
                     RDYAGRPTPLYFARRLSEKYGARVYLKREDLLHTGAHKINNAIGQVLLAKLMGKTRIIA
                     ETGAGQHGVATATAAALFGMECVIYMGEEDTIRQKLNVERMKLLGAKVVPVKSGSRTLK
                     DAIDEALRDWITNLQTTYYVIGSVVGPHPYPIIVRNFQKVIGEETKKQIPEKEGRLPDY
                     IVACVGGGSNAAGIFYPFIDSGVKLIGVEAGGEGLETGKHAASLLKGKIGYLHGSKTFV
                     LQDDWGQVQVSHSVSAGLDYSGVGPEHAYWRETGKVLYDAVTDEEALDAFIELSRLEGI
                     IPALESSHALAYLKKINIKGKVVVVNLSGRGDKDLESVLNHPYVRERIR"
     CDS             5641..5658
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      5725..5772
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5809..6264
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(join(6357..6460,1..712))
                     /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"
ORIGIN
        1 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
       61 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
      121 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
      181 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
      241 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
      301 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
      361 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
      421 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
      481 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
      541 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
      601 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
      661 tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc
      721 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt
      781 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
      841 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
      901 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
      961 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     1021 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1081 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1141 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1201 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1261 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1321 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1381 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     1441 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     1501 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     1561 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     1621 tttctcctta cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa
     1681 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     1741 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     1801 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     1861 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     1921 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     1981 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     2041 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     2101 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     2161 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     2221 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     2281 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     2341 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     2401 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     2461 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat
     2521 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
     2581 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
     2641 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
     2701 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
     2761 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     2821 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2881 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2941 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     3001 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     3061 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     3121 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     3181 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     3241 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     3301 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     3361 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     3421 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     3481 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     3541 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     3601 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     3661 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     3721 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     3781 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     3841 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     3901 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     3961 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     4021 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
     4081 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     4141 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     4201 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     4261 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
     4321 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
     4381 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
     4441 agcggcctgg tgccgcgcgg cagccatatg aaaggctact tcggtccgta cggtggtcag
     4501 tacgttccgg aaatcctgat gggtgcgctg gaagaactgg aagctgctta cgaagaaatc
     4561 atgaaagatg aatctttctg gaaagaattc aacgatctgc tgcgtgatta cgcgggtcgt
     4621 ccgaccccgc tgtacttcgc tcgtcgtctg tctgaaaaat acggtgctcg cgtttacctg
     4681 aaacgtgaag atctgctgca caccggtgcg cacaaaatca acaacgcgat cggtcaggtt
     4741 ctgctggcga aactgatggg taaaacccgt attattgctg aaaccggtgc tggccagcac
     4801 ggcgttgcga ccgctaccgc tgcggcgctg ttcggcatgg aatgcgttat ctacatgggt
     4861 gaagaagata ccatccgtca gaaactgaac gttgaacgca tgaaactgct gggtgctaaa
     4921 gttgttccgg ttaaaagcgg ttcccgtacc ctgaaagatg ctatcgatga agcgctgcgt
     4981 gattggatca ccaacctgca gaccacctat tacgttattg gtagcgttgt tggtccgcac
     5041 ccgtatccga tcatcgttcg taacttccag aaagttattg gtgaagaaac caaaaaacag
     5101 attccggaaa aagaaggtcg tctgccggat tacattgttg cgtgtgttgg tggtggtagc
     5161 aacgcggctg gtatcttcta tccgttcatc gattctggtg ttaaactgat cggtgttgaa
     5221 gcgggtggtg aaggcctgga aaccggtaaa cacgcggcta gcctgctgaa aggtaaaatt
     5281 ggctacctgc acggttctaa aaccttcgtg ctgcaagatg attggggtca ggttcaggtt
     5341 tctcattctg tgagcgcagg tctggattac tctggtgttg gtccggaaca cgcttactgg
     5401 cgtgaaaccg gtaaagttct gtacgatgct gtgaccgatg aagaagctct ggatgcgttt
     5461 attgaactga gccgtctgga aggtattatc ccagcgctgg aatcctctca cgcgctggcg
     5521 tatctgaaaa agattaacat caaaggtaaa gttgttgttg ttaacctgag cggtcgtggt
     5581 gataaagatc tggaaagcgt tctgaaccac ccgtacgttc gtgaacgtat ccgtctcgag
     5641 caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg
     5701 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     5761 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
     5821 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5881 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5941 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6001 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     6061 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6121 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     6181 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6241 acaaaatatt aacgtttaca atttcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6301 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag
     6361 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     6421 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
//