LOCUS       sequence-256057-        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     22..40
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(64..78,79..84,85..101,102..108)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    84..100
                     /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    116..132
                     /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             151..162
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             181..198
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             205..1302
                     /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     1276..1299
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1366..1386
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(2617..2823,2824..2826,2827..2946)
                     /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      2974..3068
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3112..3771
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3178..3197)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3836..3922
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4015..5097)
                     /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     5059..5078
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5098..5175)
                     /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(5259..5281)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5367..5507
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5422..5441
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5522..5539)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5693..6281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5773..5792)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6452..7243,7244..7312))
                     /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     7075..7094
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7313..7417)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct tcacctcgag
       61 aaatcataaa aaatttattt gctttgtgag cggataacaa ttataataga ttcaattgtg
      121 agcggataac aatttcacac agaattcatt aaagaggaga aattaactat gagaggatcg
      181 catcaccatc accatcacgg atctaaaatc gaagaaggta aactggtaat ctggattaac
      241 ggcgataaag gctataacgg tctcgctgaa gtcggtaaga aattcgagaa agataccgga
      301 attaaagtca ccgttgagca tccggataaa ctggaagaga aattcccaca ggttgcggca
      361 actggcgatg gccctgacat tatcttctgg gcacacgacc gctttggtgg ctacgctcaa
      421 tctggcctgt tggctgaaat caccccggac aaagcgttcc aggacaagct gtatccgttt
      481 acctgggatg ccgtacgtta caacggcaag ctgattgctt acccgatcgc tgttgaagcg
      541 ttatcgctga tttataacaa agacctgctg ccgaacccgc caaaaacctg ggaagagatc
      601 ccggcgctgg ataaagaact gaaagcgaaa ggtaagagcg cgctgatgtt caacctgcaa
      661 gaaccgtact tcacctggcc gctgattgct gctgacgggg gttatgcgtt caagtatgaa
      721 aacggcaagt acgacattaa agacgtgggc gtggataacg ctggcgcgaa agcgggtctg
      781 accttcctgg ttgacctgat taaaaacaaa cacatgaatg cagacaccga ttactccatc
      841 gcagaagctg cctttaataa aggcgaaaca gcgatgacca tcaacggccc gtgggcatgg
      901 tccaacatcg acaccagcaa agtgaattat ggtgtaacgg tactgccgac cttcaagggt
      961 caaccatcca aaccgttcgt tggcgtgctg agcgcaggta ttaacgccgc cagtccgaac
     1021 aaagagctgg caaaagagtt cctcgaaaac tatctgctga ctgatgaagg tctggaagcg
     1081 gttaataaag acaaaccgct gggtgccgta gcgctgaagt cttacgagga agagttggcg
     1141 aaagatccac gtattgccgc cactatggaa aacgcccaga aaggtgaaat catgccgaac
     1201 atcccgcaga tgtccgcttt ctggtatgcc gtgcgtactg cggtgatcaa cgccgccagc
     1261 ggtcgtcaga ctgtcgatga agccctgaaa gacgcgcaga ctaattcgag ctcgaacaac
     1321 aacaacaata acaataacaa caacctcggg atcgagggaa cgaccgaaaa cctgtatttt
     1381 cagggatccg gtagtatgtc cgcggaacga gagattcccg ccgaggacag catcaaagtg
     1441 gtctgccgat tccgaccgct gaacgacagc gaagagaagg ccggctccaa gttcgtggtc
     1501 aagttcccca acaatgtgga ggagaactgc atatccatag cgggcaaggt gtatttgttc
     1561 gacaaggtct tcaaaccgaa tgcatcccag gaaaaggtct acaatgaggc ggccaagtcc
     1621 attgttacgg atgtcctggc cgggtacaat ggaacgatat tcgcatatgg tcagacgtcc
     1681 tccggaaaaa cgcatacgat ggagggcgtg atcggggact ccgtaaaaca gggtatcata
     1741 ccacgtatcg tcaacgacat tttcaatcac atctacgcga tggaggtgaa cctggagttt
     1801 cacatcaagg tctcctacta cgagatctac atggacaaga ttcgagatct gttggacgtc
     1861 tccaaggtga accttagtgt gcacgaggat aagaaccggg tgccgtacgt caagggcgct
     1921 acggaacggt tcgtctcgtc gccggaggat gttttcgagg tgatcgagga gggcaaatcc
     1981 aatcgtcaca tcgctgtgac aaacatgaac gagcattctt cgcgatccca ctcagtattc
     2041 cttatcaatg tgaagcagga gaacctggag aaccagaaga aactatccgg caaactctac
     2101 ctggtggatt tggccggttc cgagaaggtt tccaagactg gagcggaggg aaccgttctt
     2161 gatgaagcca agaacatcaa caagtcgctg tcggccttgg gcaacgtaat ttctgccctg
     2221 gcggacggaa acaaaacgca catcccctac cgtgattcca agctaacgcg catcctgcag
     2281 gagtcgctgg gaggcaacgc acgcacaacc atcgtcatct gctgctctcc agccagtttc
     2341 aacgaatctg aaacgaagtc aacgctggac ttcggtcgta gagccaagac agtgaagaac
     2401 gtggtctgcg ttaacgagga gcttactgcc gaggaatgga agcgacgcta tgaaaaggag
     2461 aaggaaaaga acgcccgact aaagggtaag gtggagaagc tggagatcga gcttgcgcgc
     2521 tggagagcgg gtgaaactgt taaggcggag gagcaaatca acatggagga tctcatggag
     2581 gcaagcacgc ccaacctggg tagtggtagt ggatccagct ccccgaaacg ccctaagctg
     2641 ctgcgtgaat attacgattg gctggttgat aacagcttta ccccatatct ggtggtggat
     2701 gccacatacc tgggcgtgaa cgtgcccgtg gagtatgtga aagacggtca gatcgtgctg
     2761 aatctgtctg caagtgcgac cggcaacctg caactgacaa atgattttat ccagttcaac
     2821 gcccagttta agggcgtgtc tcgtgaactg tatatcccga tgggtgccgc tctggccatt
     2881 tacgctcgcg agaacggcga tggtgtgatg ttcgaaccag aagaaatcta tgacgagctg
     2941 aatattggtt aaaagcttaa ttagctgagc ttggactcct gttgatagat ccagtaatga
     3001 cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg
     3061 gtgagaatcc aagctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa
     3121 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
     3181 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
     3241 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     3301 gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg
     3361 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     3421 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     3481 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     3541 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     3601 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     3661 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     3721 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa
     3781 ggcagttatt ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat
     3841 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     3901 cgctctcctg agtaggacaa atccgccctc tagattacgt gcagtcgatg ataagctgtc
     3961 aaacatgaga attgtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     4021 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4081 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     4141 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     4201 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     4261 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     4321 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     4381 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     4441 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     4501 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     4561 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     4621 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     4681 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     4741 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     4801 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     4861 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     4921 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     4981 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     5041 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     5101 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgca
     5161 ccattcgatg gtgtcggaat ttcgggcagc gttgggtcct ggccacgggt gcgcatgatc
     5221 tagagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5281 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5341 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     5401 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     5461 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     5521 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5581 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5641 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5701 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5761 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5821 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5881 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5941 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6001 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6061 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6121 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6181 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6241 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6301 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6361 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6421 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6481 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6541 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6601 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6661 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6721 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6781 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6841 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6901 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6961 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7021 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7081 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7141 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7201 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7261 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7321 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7381 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7441 ctgacgtcta agaaaccatt att
//
