LOCUS       sequence-374269-        7355 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 7355)
  AUTHORS   Lemma LM, Varghese M, Ross TD, Thomson M, Baskaran A, Dogic Z
  TITLE     Spatio-temporal patterning of extensile active stresses in 
            microtubule-based active fluids.
  JOURNAL   PNAS Nexus. 2023 Apr 12;2(5):pgad130. doi: 
            10.1093/pnasnexus/pgad130. eCollection 2023 May.
  PUBMED    37168671
REFERENCE   2  (bases 1 to 7355)
  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"; journalName: "PNAS 
            Nexus."; date: "2023-04-12"; volume: "2(5):pgad130. doi"; pages: " 
            10.1093/pnasnexus/pgad130. eCollection 2023 May"
COMMENT     Sequence Label: K365-micro
FEATURES             Location/Qualifiers
     source          1..7355
                     /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(4242..4448,4449..4451,4452..4571)
                     /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      4599..4693
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             4737..5396
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4803..4822)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      5461..5547
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5640..6722)
                     /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     6684..6703
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6723..6800)
                     /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(6884..6906)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6992..7132
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7047..7066
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7147..7164)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7318..7355,1..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 aagggtagtg gtagtggatc cagctccccg aaacgcccta
     4261 agctgctgcg tgaatattac gattggctgg ttgataacag ctttacccca tatctggtgg
     4321 tggatgccac atacctgggc gtgaacgtgc ccgtggagta tgtgaaagac ggtcagatcg
     4381 tgctgaatct gtctgcaagt gcgaccggca acctgcaact gacaaatgat tttatccagt
     4441 tcaacgccca gtttaagggc gtgtctcgtg aactgtatat cccgatgggt gccgctctgg
     4501 ccatttacgc tcgcgagaac ggcgatggtg tgatgttcga accagaagaa atctatgacg
     4561 agctgaatat tggttaaaag cttaattagc tgagcttgga ctcctgttga tagatccagt
     4621 aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt
     4681 tattggtgag aatccaagct agcttggcga gattttcagg agctaaggaa gctaaaatgg
     4741 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     4801 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
     4861 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     4921 ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc
     4981 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     5041 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     5101 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     5161 tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     5221 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
     5281 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
     5341 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt
     5401 tttaaggcag ttattggtgc ccttaaacgc ctggggtaat gactctctag cttgaggcat
     5461 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     5521 gtgaacgctc tcctgagtag gacaaatccg ccctctagat tacgtgcagt cgatgataag
     5581 ctgtcaaaca tgagaattgt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     5641 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5701 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     5761 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     5821 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     5881 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
     5941 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     6001 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     6061 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     6121 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     6181 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     6241 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     6301 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     6361 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     6421 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     6481 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     6541 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     6601 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     6661 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     6721 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     6781 ttgcaccatt cgatggtgtc ggaatttcgg gcagcgttgg gtcctggcca cgggtgcgca
     6841 tgatctagag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     6901 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     6961 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     7021 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
     7081 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc
     7141 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7201 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7261 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7321 ccataggctc cgcccccctg acgagcatca caaaa
//
