LOCUS       sequence-161465-        5656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses monomeric streptavidin (mSA) in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5656)
  AUTHORS   Chamma I, Letellier M, Butler C, Tessier B, Lim KH, Gauthereau I, 
            Choquet D, Sibarita JB, Park S, Sainlos M, Thoumine O
  TITLE     Mapping the dynamics and nanoscale organization of synaptic adhesion
            proteins using monomeric streptavidin.
  JOURNAL   Nat Commun. 2016 Mar 16;7:10773. doi: 10.1038/ncomms10773.
  PUBMED    26979420
REFERENCE   2  (bases 1 to 5656)
  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: "Nat 
            Commun."; date: "2016-03-16"; volume: "7:10773. doi"; pages: " 
            10.1038/ncomms10773"
COMMENT     Sequence Label: pET-IG-mSA
FEATURES             Location/Qualifiers
     source          1..5656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     122..139
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    154..293
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(219..238)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     379..401
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(395..586)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1395..2477)
                     /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     2439..2458
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2478..2555)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2761..2780
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2864..2883
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2864..2882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2883..2907
                     /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             2922..2944
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2961..2990
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2991..3011
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3018..3359
                     /codon_start=1
                     /product="monomeric streptavidin/rhizavidin hybrid (Lim et 
                     al., 2013)"
                     /label=mSA
                     /translation="AEAGITGTWYNQSGSTFTVTAGADGNLTGQYENRAQGTGCQNSPY
                     TLTGRYNGTKLEWRVEWNNSTENCHSRTEWRGQYQGGAEARINTQWNLTYEGGSGPATE
                     QGQDTFTKVK"
     CDS             3384..3401
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3454..3472)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3468..3515
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3552..4007
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3639..3658)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3849..3870
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4099..4914)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4099..4914)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4822..4841
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4906..4925
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5036..5624
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5525..5544
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
       61 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      121 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
      181 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
      241 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
      301 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
      361 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
      421 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
      481 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
      541 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
      601 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
      661 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
      721 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
      781 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
      841 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
      901 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
      961 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     1021 tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct
     1081 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     1141 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     1201 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     1261 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     1321 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     1381 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1441 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     1501 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     1561 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     1621 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
     1681 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     1741 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     1801 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     1861 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     1921 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     1981 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     2041 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     2101 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     2161 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     2221 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     2281 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     2341 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     2401 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     2461 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     2521 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     2581 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     2641 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     2701 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     2761 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     2821 gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata
     2881 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
     2941 gagatataca tatggctagc caccatcacc atcatcatca tcaccatcac gaaaatctgt
     3001 attttcaaag cggatccgcg gaagcgggta tcaccggcac gtggtacaac cagtctggtt
     3061 ctaccttcac cgttaccgcg ggtgcggacg gtaacctgac cggtcagtac gaaaaccgtg
     3121 cgcagggcac tggttgccag aactctccgt acaccctgac cggtcgttac aacggtacca
     3181 aactggaatg gcgtgttgaa tggaacaact ctaccgaaaa ctgccactct cgtaccgaat
     3241 ggcgtggtca gtaccagggt ggtgcggaag cgcgtatcaa cacccagtgg aacctgacct
     3301 acgaaggtgg ttctggtccg gcgaccgaac agggtcagga caccttcacc aaagttaaac
     3361 cgtctgcggc gtcttaactc gagcaccacc accaccacca ctgagatccg gctgctaaca
     3421 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     3481 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
     3541 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3601 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3661 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3721 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3781 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3841 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3901 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3961 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     4021 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4081 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     4141 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     4201 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     4261 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     4321 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     4381 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     4441 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     4501 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     4561 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     4621 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     4681 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     4741 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     4801 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     4861 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc
     4921 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac
     4981 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5041 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5101 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5161 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     5221 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5281 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5341 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5401 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5461 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5521 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5581 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5641 cctttttacg gttcct
//
