LOCUS       sequence-190912-        7287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Histag-SUMO-FtsZ for purification in E coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7287)
  AUTHORS   Yang X, Lyu Z, Miguel A, McQuillen R, Huang KC, Xiao J
  TITLE     GTPase activity-coupled treadmilling of the bacterial tubulin FtsZ 
            organizes septal cell wall synthesis.
  JOURNAL   Science. 2017 Feb 17;355(6326):744-747. doi: 
            10.1126/science.aak9995.
  PUBMED    28209899
REFERENCE   2  (bases 1 to 7287)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2017-02-17- 17"; volume: "355"; 
            issue: "6326"; pages: "744-747"
COMMENT     Sequence Label: pRM006
FEATURES             Location/Qualifiers
     source          1..7287
                     /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             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             130..420
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     primer_bind     complement(1658..1676)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1672..1719
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1907..1924)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1907..1923)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1916..1938)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2102..2119)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2272..2358
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2450..2477
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2496..2587
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2588..2656,2657..3448)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2806..2825)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3490..4003)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3685..3706)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3897..3916
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4114..4702
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4603..4622
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4856..4873
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4895..4914)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5055..5077
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5071..5262)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5819..6901)
                     /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     6863..6882
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6902..6979)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7185..7204
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg cgtggttctc accaccacca ccaccatatg
      121 gctagcggat cggactcaga agtcaatcaa gaagctaagc cagaggtcaa gccagaagtc
      181 aagcctgaga ctcacatcaa tttaaaggtg tccgatggat cttcagagat cttcttcaag
      241 atcaaaaaga ccactccttt aagaaggctg atggaagcgt tcgctaaaag acagggtaag
      301 gaaatggact ccttaagatt cttgtacgac ggtattagaa ttcaagctga tcagacccct
      361 gaagatttgg acatggagga taacgatatt attgaggctc acagagaaca gattggtggt
      421 atgtttgaac caatggaact taccaatgac gcggtgatta aagtcatcgg cgtcggcggc
      481 ggcggcggta atgctgttga acacatggtg cgcgagcgca ttgaaggtgt tgaattcttc
      541 gcggtaaata ccgatgcaca agcgctgcgt aaaacagcgg ttggacagac gattcaaatc
      601 ggtagcggta tcaccaaagg actgggcgct ggcgctaatc cagaagttgg ccgcaatgcg
      661 gctgatgagg atcgcgatgc attgcgtgcg gcgctggaag gtgcagacat ggtctttatt
      721 gctgcgggta tgggtggtgg taccggtaca ggtgcagcac cagtcgtcgc tgaagtggca
      781 aaagatttgg gtatcctgac cgttgctgtc gtcactaagc ctttcaactt tgaaggcaag
      841 aagcgtatgg cattcgcgga gcaggggatc actgaactgt ccaagcatgt ggactctctg
      901 atcactatcc cgaacgacaa actgctgaaa gttctgggcc gcggtatctc cctgctggat
      961 gcgtttggcg cagcgaacga tgtactgaaa ggcgctgtgc aaggtatcgc tgaactgatt
     1021 actcgtccgg gtttgatgaa cgtggacttt gcagacgtac gcaccgtaat gtctgagatg
     1081 ggctacgcaa tgatgggttc tggcgtggcg agcggtgaag accgtgcgga agaagctgct
     1141 gaaatggcta tctcttctcc gctgctggaa gatatcgacc tgtctggcgc gcgcggcgtg
     1201 ctggttaaca tcacggcggg cttcgatctg cgtctggatg agttcgaaac ggtaggtaac
     1261 accatccgtg catttgcttc cgacaacgcg actgtggtta tcggtacttc tcttgacccg
     1321 gatatgaatg acgagctgcg cgtaaccgtt gttgcgacag gtatcggcat ggacaaacgt
     1381 cctgaaatca ctctggtgac caataagcag gttcagcagc cagtgatgga tcgctaccag
     1441 cagcatggga tggctccgct gacccaggag cagaagccgg ttgctaaagt cgtgaatgac
     1501 aatgcgccgc aaactgcgaa agagccggat tatctggata tcccagcatt cctgcgtaag
     1561 caagctgatt aagcggccgc actcgagccc gggtgactgc aggaagggga tccggctgct
     1621 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     1681 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     1741 ggatatcccg caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca
     1801 gggtgacggt gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac
     1861 tgcgttagca atttaactgt gataaactac cgcattaaag cttggcactg gccgtcgttt
     1921 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     1981 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     2041 tgcgcagcct gaatggcgaa tggcagcttg gctgttttgg cggatgagat aagattttca
     2101 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
     2161 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
     2221 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     2281 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     2341 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     2401 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
     2461 ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct aaatacattc
     2521 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2581 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2641 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2701 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2761 tcgccccgaa gaacgtttcc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2821 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2881 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2941 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3001 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3061 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3121 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3181 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3241 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3301 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3361 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3421 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3481 gattgattta ccccggttga taatcagaaa agccccaaaa acaggaagat tgtataagca
     3541 aatatttaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3601 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3661 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3721 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3781 tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3841 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     3901 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     3961 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtaaaagga tctaggtgaa
     4021 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4081 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4141 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4201 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4261 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4321 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4381 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4441 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4501 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4561 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4621 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4681 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4741 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4801 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4861 gtcagtgagc gaggaagcta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4921 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     4981 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5041 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5101 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtgc agcgattcac agatgtctgc
     5161 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     5221 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
     5281 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
     5341 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
     5401 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagccga acgccagcaa
     5461 gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     5521 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     5581 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     5641 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     5701 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     5761 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     5821 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5881 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     5941 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     6001 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     6061 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     6121 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     6181 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     6241 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     6301 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     6361 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     6421 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     6481 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     6541 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     6601 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     6661 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     6721 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     6781 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     6841 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     6901 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     6961 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     7021 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     7081 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     7141 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     7201 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     7261 aggatcgaga tctcgatccc gcgaaat
//
