LOCUS       sequence-197129-        6772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  production of gamma-tubulin in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6772)
  AUTHORS   Alvarado-Kristensson M, Rodriguez MJ, Silio V, Valpuesta JM, Carrera
            AC
  TITLE     SADB phosphorylation of gamma-tubulin regulates centrosome 
            duplication.
  JOURNAL   Nat Cell Biol. 2009 Sep;11(9):1081-92. doi: 10.1038/ncb1921. Epub 
            2009 Aug 2.
  PUBMED    19648910
REFERENCE   2  (bases 1 to 6772)
  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: "10.1038/ncb1921"; 
            journalName: "Nat Cell Biol"; date: "2009-09"; volume: "11"; issue: 
            "9"; pages: "1081-92"
COMMENT     Sequence Label: pET21 gamma-tubulin
FEATURES             Location/Qualifiers
     source          1..6772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             88..120
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             1504..1521
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1574..1592)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1588..1635
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1672..2127
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1759..1778)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1969..1990
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2153..2257
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2258..2326,2327..3118)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2476..2495)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3289..3877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3778..3797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4031..4048
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4063..4202
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4128..4147)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4288..4310
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4304..4495)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5304..6386)
                     /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     6348..6367
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6387..6464)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6670..6689
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg gctagcatga ctggtggaca gcaaatgggt
      121 cggatccgaa ttcgtatgcc gagggaaatc atcaccctac agttgggcca gtgcggcaat
      181 cagattgggt tcgagttctg gaaacagctg tgcgccgagc atggtatcag ccccgagggc
      241 atcgtggagg agttcgccac cgagggcact gaccacaagg acgtcttttt ctaccaggca
      301 gacgatgagc actacatccc ccgggccgtg ctgctggact tggaaccccg ggtgatccac
      361 tccatcctca actcccccta tgccaagctc tacaacccag agaacatcta cctgtcggaa
      421 catggaggag gagctggcaa caactgggcc agcggattct cccagggaga aaagatccat
      481 gaggacattt ttgacatcat agaccgggag gcagatggta gtgacagtct agagggcttt
      541 gtgctgtgtc actccattgc tggggggaca ggctctggac tgggttccta cctcttagaa
      601 cggctgaatg acaggtatcc taagaagctg gtgcagacat actcagtgtt tcccaaccag
      661 gacgagatga gcgatgtggt ggtccagcct tacaattcac tcctcacact caagaggctg
      721 acgcagaatg cagactgtgt ggtggtgctg gacaacacag ccctgaaccg gattgccaca
      781 gaccgcctgc acatccagaa cccatccttc tcccagatca accagctggt gtctaccatc
      841 atgtcagcca gcaccaccac cctgcgctac cctggctaca tgaacaatga cctcatcggc
      901 ctcatcgcct cgctcattcc caccccacgg ctccacttcc tcatgaccgg ctacacccct
      961 ctcactacgg accagtcagt ggccagcgtg aggaagacca cggtcctgga tgtcatgagg
     1021 cggctgctgc agcccaagaa cgtgatggtg tccacaggcc gagaccgcca gaccaaccac
     1081 tgctacatcg ccatcctcaa catcatccag ggagaggtgg accccaccca ggtccacaag
     1141 agcttgcaga ggatccggga acgcaagttg gccaacttca tcccgtgggg ccccgccagc
     1201 atccaggtgg ccctgtcgag gaagtctccc tacctgccct cggcccaccg ggtcagcggg
     1261 ctcatgatgg ccaaccacac cagcatctcc tcgctcttcg agagaacctg tcgccagtat
     1321 gacaagctgc gtaagcggga ggccttcctg gagcagttcc gcaaggagga catgttcaag
     1381 gacaactttg atgagatgga cacatccagg gagattgtgc agcagctcat cgatgagtac
     1441 catgcggcca cacggccaga ctacatctcc tggggcaccc agaagcttgc ggccgcactc
     1501 gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag
     1561 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     1621 ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct
     1681 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     1741 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     1801 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     1861 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     1921 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     1981 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2041 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2101 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     2161 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2221 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     2281 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     2341 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     2401 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     2461 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     2521 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     2581 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     2641 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     2701 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2761 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     2821 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     2881 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2941 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3001 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3061 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3121 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3181 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3241 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3301 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3361 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3421 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3481 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3541 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3601 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3661 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3721 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3781 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3841 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3901 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3961 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4021 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     4081 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     4141 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     4201 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4261 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4321 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     4381 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     4441 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     4501 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     4561 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     4621 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     4681 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     4741 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     4801 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     4861 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     4921 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga
     4981 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     5041 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     5101 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     5161 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     5221 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     5281 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5341 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     5401 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     5461 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     5521 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     5581 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     5641 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     5701 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     5761 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     5821 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     5881 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     5941 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     6001 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     6061 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     6121 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     6181 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     6241 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     6301 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     6361 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     6421 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     6481 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     6541 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     6601 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     6661 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     6721 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa at
//
