LOCUS       sequence-252167-        5273 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BioBrick pET vector for protein overexpression in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5273)
  AUTHORS   Du L, Gao R, Forster AC
  TITLE     Engineering multigene expression in vitro and in vivo with small 
            terminators for T7 RNA polymerase.
  JOURNAL   Biotechnol Bioeng. 2009 Dec 15;104(6):1189-96. doi: 
            10.1002/bit.22491.
  PUBMED    19650080
REFERENCE   2  (bases 1 to 5273)
  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"; doi: "10.1002/bit"; 
            journalName: "Biotechnol Bioeng"; date: "2009-12-15- 15"; volume: 
            "104"; issue: "6"; pages: "1189-96"
COMMENT     Sequence Label: BioBrick pET-24a(+)
FEATURES             Location/Qualifiers
     source          1..5273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..22
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     primer_bind     37..56
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        37..55
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    56..80
                     /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             95..117
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(207..225)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      221..268
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    283..303
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     rep_origin      322..777
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(409..428)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     619..640
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(869..1684)
                     /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(869..1684)
                     /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     1592..1611
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1676..1695
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1806..2392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2293..2312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2546..2563
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2578..2717
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2643..2662)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2803..2825
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2819..3010)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3819..4901)
                     /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     4863..4882
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4902..4979)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5185..5204
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gaattcgcgg ccgcttctag agcgatcccg cgaaattaat acgactcact ataggggaat
       61 tgtgagcgga taacaattcc ccactagaaa taattttgtt taactttaag aaggagatat
      121 acatatgcac cactcgagtt atctgttagt ttttttctac tagactgcta acaaagcccg
      181 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
      241 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actactagta gcggccgctg
      301 cagtccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
      361 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
      421 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
      481 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
      541 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
      601 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
      661 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
      721 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
      781 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      841 aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact
      901 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
      961 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     1021 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     1081 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca
     1141 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     1201 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     1261 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     1321 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     1381 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     1441 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     1501 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     1561 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     1621 cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc
     1681 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa
     1741 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1801 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1861 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttca aggtaactgg
     1921 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     1981 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2041 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2101 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2161 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2221 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2281 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2341 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2401 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2461 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2521 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     2581 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     2641 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     2701 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2761 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     2821 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     2881 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     2941 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     3001 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     3061 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     3121 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     3181 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     3241 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     3301 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     3361 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     3421 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg
     3481 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     3541 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     3601 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     3661 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     3721 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     3781 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3841 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3901 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     3961 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     4021 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     4081 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     4141 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     4201 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     4261 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     4321 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     4381 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     4441 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     4501 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     4561 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     4621 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     4681 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     4741 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     4801 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     4861 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     4921 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     4981 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     5041 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     5101 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     5161 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     5221 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tct
//
