LOCUS       sequence-233782-        7463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Component of optically-induced motor dimerization system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7463)
  AUTHORS   Ross TD, Lee HJ, Qu Z, Banks RA, Phillips R, Thomson M
  TITLE     Controlling organization and forces in active matter through 
            optically defined boundaries.
  JOURNAL   Nature. 2019 Aug;572(7768):224-229. doi: 10.1038/s41586-019-1447-1. 
            Epub 2019 Aug 7.
  PUBMED    31391558
REFERENCE   2  (bases 1 to 7463)
  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/s41586-019-1447-1"; journalName: "Nature"; date: "2019-08";
            volume: "572"; issue: "7768"; pages: "224-229"
COMMENT     Sequence Label: K401-Micro
FEATURES             Location/Qualifiers
     source          1..7463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(722..1513,1514..1582))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1345..1364
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1583..1687)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1755..1773
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1797..1811,1812..1817,1818..1834,1835..1841)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    1817..1833
                     /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)."
     protein_bind    1849..1865
                     /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             1884..1895
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1914..1931
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1938..3035
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     3009..3032
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             3099..3119
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(4350..4556,4557..4559,4560..4679)
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein and with the mutation R74Q to reduce the 
                     affinity for iLID"
                     /label=SspB micro
                     /note="dimerizes with iLID at moderate affinity to create 
                     iLID micro (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNAQFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
     terminator      4707..4801
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             4845..5504
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4911..4930)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      5569..5655
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5748..6830)
                     /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     6792..6811
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6831..6908)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(6992..7014)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7100..7240
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7155..7174
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7255..7272)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7426..7463,1..551))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
       61 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
      121 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
      181 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
      241 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
      301 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
      361 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
      421 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
      481 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
      541 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
      601 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
      661 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
      721 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
      781 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
      841 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
      901 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
      961 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     1021 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     1081 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     1141 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     1201 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     1261 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     1321 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     1381 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     1441 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     1501 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     1561 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     1621 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     1681 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     1741 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcacctc gagaaatcat
     1801 aaaaaattta tttgctttgt gagcggataa caattataat agattcaatt gtgagcggat
     1861 aacaatttca cacagaattc attaaagagg agaaattaac tatgagagga tcgcatcacc
     1921 atcaccatca cggatctaaa atcgaagaag gtaaactggt aatctggatt aacggcgata
     1981 aaggctataa cggtctcgct gaagtcggta agaaattcga gaaagatacc ggaattaaag
     2041 tcaccgttga gcatccggat aaactggaag agaaattccc acaggttgcg gcaactggcg
     2101 atggccctga cattatcttc tgggcacacg accgctttgg tggctacgct caatctggcc
     2161 tgttggctga aatcaccccg gacaaagcgt tccaggacaa gctgtatccg tttacctggg
     2221 atgccgtacg ttacaacggc aagctgattg cttacccgat cgctgttgaa gcgttatcgc
     2281 tgatttataa caaagacctg ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc
     2341 tggataaaga actgaaagcg aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt
     2401 acttcacctg gccgctgatt gctgctgacg ggggttatgc gttcaagtat gaaaacggca
     2461 agtacgacat taaagacgtg ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc
     2521 tggttgacct gattaaaaac aaacacatga atgcagacac cgattactcc atcgcagaag
     2581 ctgcctttaa taaaggcgaa acagcgatga ccatcaacgg cccgtgggca tggtccaaca
     2641 tcgacaccag caaagtgaat tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat
     2701 ccaaaccgtt cgttggcgtg ctgagcgcag gtattaacgc cgccagtccg aacaaagagc
     2761 tggcaaaaga gttcctcgaa aactatctgc tgactgatga aggtctggaa gcggttaata
     2821 aagacaaacc gctgggtgcc gtagcgctga agtcttacga ggaagagttg gcgaaagatc
     2881 cacgtattgc cgccactatg gaaaacgccc agaaaggtga aatcatgccg aacatcccgc
     2941 agatgtccgc tttctggtat gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc
     3001 agactgtcga tgaagccctg aaagacgcgc agactaattc gagctcgaac aacaacaaca
     3061 ataacaataa caacaacctc gggatcgagg gaacgaccga aaacctgtat tttcagggat
     3121 ccggtagtat gtccgcggaa cgagagattc ccgccgagga cagcatcaaa gtggtctgcc
     3181 gattccgacc gctgaacgac agcgaagaga aggccggctc caagttcgtg gtcaagttcc
     3241 ccaacaatgt ggaggagaac tgcatatcca tagcgggcaa ggtgtatttg ttcgacaagg
     3301 tcttcaaacc gaatgcatcc caggaaaagg tctacaatga ggcggccaag tccattgtta
     3361 cggatgtcct ggccgggtac aatggaacga tattcgcata tggtcagacg tcctccggaa
     3421 aaacgcatac gatggagggc gtgatcgggg actccgtaaa acagggtatc ataccacgta
     3481 tcgtcaacga cattttcaat cacatctacg cgatggaggt gaacctggag tttcacatca
     3541 aggtctccta ctacgagatc tacatggaca agattcgaga tctgttggac gtctccaagg
     3601 tgaaccttag tgtgcacgag gataagaacc gggtgccgta cgtcaagggc gctacggaac
     3661 ggttcgtctc gtcgccggag gatgttttcg aggtgatcga ggagggcaaa tccaatcgtc
     3721 acatcgctgt gacaaacatg aacgagcatt cttcgcgatc ccactcagta ttccttatca
     3781 atgtgaagca ggagaacctg gagaaccaga agaaactatc cggcaaactc tacctggtgg
     3841 atttggccgg ttccgagaag gtttccaaga ctggagcgga gggaaccgtt cttgatgaag
     3901 ccaagaacat caacaagtcg ctgtcggcct tgggcaacgt aatttctgcc ctggcggacg
     3961 gaaacaaaac gcacatcccc taccgtgatt ccaagctaac gcgcatcctg caggagtcgc
     4021 tgggaggcaa cgcacgcaca accatcgtca tctgctgctc tccagccagt ttcaacgaat
     4081 ctgaaacgaa gtcaacgctg gacttcggtc gtagagccaa gacagtgaag aacgtggtct
     4141 gcgttaacga ggagcttact gccgaggaat ggaagcgacg ctatgaaaag gagaaggaaa
     4201 agaacgcccg actaaagggt aaggtggaga agctggagat cgagcttgcg cgctggagag
     4261 cgggtgaaac tgttaaggcg gaggagcaaa tcaacatgga ggatctcatg gaggcaagca
     4321 cgcccaacct gggtagtggt agtggatcca gctccccgaa acgccctaag ctgctgcgtg
     4381 aatattacga ttggctggtt gataacagct ttaccccata tctggtggtg gatgccacat
     4441 acctgggcgt gaacgtgccc gtggagtatg tgaaagacgg tcagatcgtg ctgaatctgt
     4501 ctgcaagtgc gaccggcaac ctgcaactga caaatgattt tatccagttc aacgcccagt
     4561 ttaagggcgt gtctcgtgaa ctgtatatcc cgatgggtgc cgctctggcc atttacgctc
     4621 gcgagaacgg cgatggtgtg atgttcgaac cagaagaaat ctatgacgag ctgaatattg
     4681 gttaaaagct taattagctg agcttggact cctgttgata gatccagtaa tgacctcaga
     4741 actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa
     4801 tccaagctag cttggcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca
     4861 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
     4921 agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa
     4981 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
     5041 tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg gtgatatggg
     5101 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct
     5161 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
     5221 gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct
     5281 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
     5341 tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc
     5401 cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta
     5461 atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt
     5521 attggtgccc ttaaacgcct ggggtaatga ctctctagct tgaggcatca aataaaacga
     5581 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     5641 ctgagtagga caaatccgcc ctctagatta cgtgcagtcg atgataagct gtcaaacatg
     5701 agaattgtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     5761 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5821 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     5881 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     5941 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     6001 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     6061 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     6121 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     6181 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     6241 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     6301 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     6361 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     6421 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     6481 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     6541 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     6601 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     6661 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     6721 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     6781 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     6841 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcaccattcg
     6901 atggtgtcgg aatttcgggc agcgttgggt cctggccacg ggtgcgcatg atctagagct
     6961 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     7021 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     7081 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     7141 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     7201 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     7261 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7321 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7381 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7441 cccccctgac gagcatcaca aaa
//
