LOCUS       sequence-224462-        6158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  surface-displayed Nb2, used in composability experiments; medium 
            copy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6158)
  AUTHORS   Glass DS, Riedel-Kruse IH
  TITLE     A Synthetic Bacterial Cell-Cell Adhesion Toolbox for Programming 
            Multicellular Morphologies and Patterns.
  JOURNAL   Cell. 2018 Jul 26;174(3):649-658.e16. doi: 
            10.1016/j.cell.2018.06.041. Epub 2018 Jul 19.
  PUBMED    30033369
REFERENCE   2  (bases 1 to 6158)
  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: 
            "10.1016/j.cell.2018.06.041"; journalName: "Cell"; date: 
            "2018-07-26- 26"; volume: "174"; issue: "3"; pages: "649-658.e16"
COMMENT     Sequence Label: pDSG320
FEATURES             Location/Qualifiers
     source          1..6158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             45..56
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             63..686
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(136..155)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      706..777
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      793..820
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    835..853
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    860..878
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             897..908
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      3289..3360
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3376..3403
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3410..3430
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3431..3488
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     CDS             complement(3646..4461)
                     /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(3646..4461)
                     /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     4369..4388
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4453..4472
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(5056..5601)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(5136..5155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6069..6087
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(6091..6134)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    6137..6158
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tggtgcaaaa cctttgcggt atggcatgat agcgcctact agagaaagag gagaaatact
       61 agatgtccag attagataaa agtaaagtga ttaacagcgc attagagctg cttaatgagg
      121 tcggaatcga aggtttaaca acccgtaaac tcgcccagaa gctaggtgta gagcagccta
      181 cattgtattg gcatgtaaaa aataagcggg ctttgctcga cgccttagcc attgagatgt
      241 tagataggca ccatactcac ttttgccctt tagaagggga aagctggcaa gattttttac
      301 gtaataacgc taaaagtttt agatgtgctt tactaagtca tcgcgatgga gcaaaagtac
      361 atttaggtac acggcctaca gaaaaacagt atgaaactct cgaaaatcaa ttagcctttt
      421 tatgccaaca aggtttttca ctagagaatg cattatatgc actcagcgct gtggggcatt
      481 ttactttagg ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa gaaagggaaa
      541 cacctactac tgatagtatg ccgccattat tacgacaagc tatcgaatta tttgatcacc
      601 aaggtgcaga gccagccttc ttattcggcc ttgaattgat catatgcgga ttagaaaaac
      661 aacttaaatg tgaaagtggg tcctaataat actagagcca ggcatcaaat aaaacgaaag
      721 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
      781 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatatact agagtcccta
      841 tcagtgatag agattgacat ccctatcagt gatagagata ctgagcacta ctagagaaag
      901 aggagaaata ctagatgatt actcatggtt gttatacccg gacccggcac aagcataagc
      961 taaaaaaaac attgattatg cttagtgctg gtttaggatt gtttttttat gttaatcaga
     1021 actcatttgc aaatggtgaa aattatttta aattgggttc ggattcaaaa ctgttaactc
     1081 atgatagcta tcagaatcgc cttttttata cgttgaaaac tggtgaaact gttgccgatc
     1141 tttctaaatc gcaagatatt aatttatcga cgatttggtc gttgaataag catttataca
     1201 gttctgaaag cgaaatgatg aaggccgcgc ctggtcagca gatcattttg ccactcaaaa
     1261 aacttccctt tgaatacagt gcactaccac ttttaggttc ggcacctctt gttgctgcgg
     1321 gtggtgttgc tggtcacacg aataaactga ctaaaatgtc cccggacgtg accaaaagca
     1381 acatgaccga tgacaaggca ttaaattatg cggcacaaca ggcggcgagt ctcggtagcc
     1441 agcttcagtc gcgatctctg aacggcgatt acgcgaaaga taccgctctt ggtatcgctg
     1501 gtaaccaggc ttcgtcacag ttgcaggcct ggttacaaca ttatggaacg gcagaggtta
     1561 atctgcaaag tggtaataac tttgacggta gttcactgga cttcttatta ccgttctatg
     1621 attccgaaaa aatgctggca tttggtcagg tcggagcgcg ttacattgac tcccgcttta
     1681 cggcaaattt aggtgcgggt cagcgttttt tccttcctgc aaacatgttg ggctataacg
     1741 tcttcattga tcaggatttt tctggtgata atacccgttt aggtattggt ggcgaatact
     1801 ggcgagacta tttcaaaagt agcgttaacg gctatttccg catgagcggc tggcatgagt
     1861 catacaataa gaaagactat gatgagcgcc cagcaaatgg cttcgatatc cgttttaatg
     1921 gctatctacc gtcatatccg gcattaggcg ccaagctgat atatgagcag tattatggtg
     1981 ataatgttgc tttgtttaat tctgataagc tgcaatcgaa tcctggtgcg gcgaccgttg
     2041 gtgtaaacta tactccgatt cctctggtga cgatggggat cgattaccgt catggtacgg
     2101 gtaatgaaaa tgatctcctt tactcaatgc agttccgtta tcagtttgat aaatcgtggt
     2161 ctcagcaaat tgaaccacag tatgttaacg agttaagaac attatcaggc agccgttacg
     2221 atctggttca gcgtaataac aatattattc tggagtacaa gaagcaggat attctttctc
     2281 tgaatattcc gcatgatatt aatggtactg aacacagtac gcagaagatt cagttgatcg
     2341 ttaagagcaa atacggtctg gatcgtatcg tctgggatga tagtgcatta cgcagtcagg
     2401 gcggtcagat tcagcatagc ggaagccaaa gcgcacaaga ctaccaggct attttgcctg
     2461 cttatgtgca aggtggcagc aatatttata aagtgacggc tcgcgcctat gaccgtaatg
     2521 gcaatagctc taacaatgta cagcttacta ttaccgttct gtcgaatggt caagttgtcg
     2581 accaggttgg ggtaacggac tttacggcgg ataagacttc ggctaaagcg gataacgccg
     2641 ataccattac ttataccgcg acggtgaaaa agaatggggt agctcaggct aatgtccctg
     2701 tttcatttaa tattgtttca ggaactgcaa ctcttggggc aaatagtgcc aaaacggatg
     2761 ctaacggtaa ggcaaccgta acgttgaagt cgagtacgcc aggacaggtc gtcgtgtctg
     2821 ctaaaaccgc ggagatgact tcagcactta atgccagtgc ggttatattt tttgatggtg
     2881 cgactagaca agggcagctt gttgagagcg gggggggaag cgtacaagcc gggggatcgc
     2941 tgcgcttgtc gtgtgcagct tccggtattg attccagctc atactgtatg ggttggtttc
     3001 gtcagcgtcc gggaaaggag cgcgagggtg tcgcacgtat caacgggttg ggcggcgtta
     3061 aaacagctta cgctgactct gttaaagatc gtttcaccat tagtcgtgat aatgcagaaa
     3121 atactgttta tttgcaaatg aactccttga aacccgaaga tacagctatt tattattgtg
     3181 ctgccaagtt tagtccaggt tattgcggag gatcgtggag caatttcgga tactggggtc
     3241 aaggaacgca agtgaccgta tcttcgtaat aatactagag ccaggcatca aataaaacga
     3301 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     3361 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatat actagtagcg
     3421 gccgctgcag tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt tctttaaaac
     3481 cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac tgactcgctg cgctcggtcg
     3541 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatctcgagt cccgtcaagt
     3601 cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc
     3661 gagcatcaaa tgaaactgca atttattcat atcaggatta tcgataccat atttttgaaa
     3721 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     3781 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     3841 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     3901 tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     3961 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     4021 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     4081 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     4141 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     4201 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     4261 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     4321 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     4381 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctgg agcaagacgt
     4441 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     4501 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     4561 aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct
     4621 tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac
     4681 ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat
     4741 tcaggcctgg tatgagtcag caacaccttc ttcacgaggc agacctcagc gctagcggag
     4801 tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca
     4861 ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt
     4921 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg
     4981 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc
     5041 cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg
     5101 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5161 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt
     5221 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca
     5281 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact
     5341 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta
     5401 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca
     5461 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag
     5521 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg
     5581 cgcagaccaa aacgatctca agaagatcat cttattaagg ggtctgacgc tcagtggaac
     5641 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5701 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5761 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5821 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5881 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5941 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6001 atccagtcta ttccatggtg ccacctgacg tctaagaaac cattattatc atgacattaa
     6061 cctataaaaa taggcgtatc acgaggcaga atttcagata aaaaaaatcc ttagctttcg
     6121 ctaaggatga tttctggaat tcgcggccgc ttctagag
//
