LOCUS       sequence-426876-        5792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  surface-displayed Ag2, used in composability experiments; medium 
            copy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5792)
  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 5792)
  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: pDSG318
FEATURES             Location/Qualifiers
     source          1..5792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             11..22
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             29..652
                     /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(102..121)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      672..743
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      759..786
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    801..819
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    826..844
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             863..874
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2889..2960
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2976..3003
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3010..3030
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3031..3088
                     /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(3246..4061)
                     /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(3246..4061)
                     /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     3969..3988
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4053..4072
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(4656..5201)
                     /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(4736..4755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5669..5687
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5691..5734)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5737..5758
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 cctactagag aaagaggaga aatactagat gtccagatta gataaaagta aagtgattaa
       61 cagcgcatta gagctgctta atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc
      121 ccagaagcta ggtgtagagc agcctacatt gtattggcat gtaaaaaata agcgggcttt
      181 gctcgacgcc ttagccattg agatgttaga taggcaccat actcactttt gccctttaga
      241 aggggaaagc tggcaagatt ttttacgtaa taacgctaaa agttttagat gtgctttact
      301 aagtcatcgc gatggagcaa aagtacattt aggtacacgg cctacagaaa aacagtatga
      361 aactctcgaa aatcaattag cctttttatg ccaacaaggt ttttcactag agaatgcatt
      421 atatgcactc agcgctgtgg ggcattttac tttaggttgc gtattggaag atcaagagca
      481 tcaagtcgct aaagaagaaa gggaaacacc tactactgat agtatgccgc cattattacg
      541 acaagctatc gaattatttg atcaccaagg tgcagagcca gccttcttat tcggccttga
      601 attgatcata tgcggattag aaaaacaact taaatgtgaa agtgggtcct aataatacta
      661 gagccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
      721 gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt
      781 tctgcgttta tatactagag tccctatcag tgatagagat tgacatccct atcagtgata
      841 gagatactga gcactactag agaaagagga gaaatactag atgattactc atggttgtta
      901 tacccggacc cggcacaagc ataagctaaa aaaaacattg attatgctta gtgctggttt
      961 aggattgttt ttttatgtta atcagaactc atttgcaaat ggtgaaaatt attttaaatt
     1021 gggttcggat tcaaaactgt taactcatga tagctatcag aatcgccttt tttatacgtt
     1081 gaaaactggt gaaactgttg ccgatctttc taaatcgcaa gatattaatt tatcgacgat
     1141 ttggtcgttg aataagcatt tatacagttc tgaaagcgaa atgatgaagg ccgcgcctgg
     1201 tcagcagatc attttgccac tcaaaaaact tccctttgaa tacagtgcac taccactttt
     1261 aggttcggca cctcttgttg ctgcgggtgg tgttgctggt cacacgaata aactgactaa
     1321 aatgtccccg gacgtgacca aaagcaacat gaccgatgac aaggcattaa attatgcggc
     1381 acaacaggcg gcgagtctcg gtagccagct tcagtcgcga tctctgaacg gcgattacgc
     1441 gaaagatacc gctcttggta tcgctggtaa ccaggcttcg tcacagttgc aggcctggtt
     1501 acaacattat ggaacggcag aggttaatct gcaaagtggt aataactttg acggtagttc
     1561 actggacttc ttattaccgt tctatgattc cgaaaaaatg ctggcatttg gtcaggtcgg
     1621 agcgcgttac attgactccc gctttacggc aaatttaggt gcgggtcagc gttttttcct
     1681 tcctgcaaac atgttgggct ataacgtctt cattgatcag gatttttctg gtgataatac
     1741 ccgtttaggt attggtggcg aatactggcg agactatttc aaaagtagcg ttaacggcta
     1801 tttccgcatg agcggctggc atgagtcata caataagaaa gactatgatg agcgcccagc
     1861 aaatggcttc gatatccgtt ttaatggcta tctaccgtca tatccggcat taggcgccaa
     1921 gctgatatat gagcagtatt atggtgataa tgttgctttg tttaattctg ataagctgca
     1981 atcgaatcct ggtgcggcga ccgttggtgt aaactatact ccgattcctc tggtgacgat
     2041 ggggatcgat taccgtcatg gtacgggtaa tgaaaatgat ctcctttact caatgcagtt
     2101 ccgttatcag tttgataaat cgtggtctca gcaaattgaa ccacagtatg ttaacgagtt
     2161 aagaacatta tcaggcagcc gttacgatct ggttcagcgt aataacaata ttattctgga
     2221 gtacaagaag caggatattc tttctctgaa tattccgcat gatattaatg gtactgaaca
     2281 cagtacgcag aagattcagt tgatcgttaa gagcaaatac ggtctggatc gtatcgtctg
     2341 ggatgatagt gcattacgca gtcagggcgg tcagattcag catagcggaa gccaaagcgc
     2401 acaagactac caggctattt tgcctgctta tgtgcaaggt ggcagcaata tttataaagt
     2461 gacggctcgc gcctatgacc gtaatggcaa tagctctaac aatgtacagc ttactattac
     2521 cgttctgtcg aatggtcaag ttgtcgacca ggttggggta acggacttta cggcggataa
     2581 gacttcggct aaagcggata acgccgatac cattacttat accgcgacgg tgaaaaagaa
     2641 tggggtagct caggctaatg tccctgtttc atttaatatt gtttcaggaa ctgcaactct
     2701 tggggcaaat agtgccaaaa cggatgctaa cggtaaggca accgtaacgt tgaagtcgag
     2761 tacgccagga caggtcgtcg tgtctgctaa aaccgcggag atgacttcag cacttaatgc
     2821 cagtgcggtt atattttttg atggtgcgac tagagaacca gaggcttaat aatactagag
     2881 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     2941 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
     3001 gcgtttatat actagtagcg gccgctgcag tccggcaaaa aagggcaagg tgtcaccacc
     3061 ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac
     3121 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3181 aatctcgagt cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat
     3241 tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     3301 tcgataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     3361 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     3421 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     3481 acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca
     3541 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     3601 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     3661 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     3721 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     3781 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     3841 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     3901 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     3961 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     4021 cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     4081 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     4141 catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg
     4201 aaggatcaga tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc
     4261 aaaatcacca actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc
     4321 tggctggatg atggggcgat tcaggcctgg tatgagtcag caacaccttc ttcacgaggc
     4381 agacctcagc gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca
     4441 gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg
     4501 atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
     4561 gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact
     4621 taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac
     4681 aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga
     4741 taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
     4801 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
     4861 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg
     4921 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
     4981 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
     5041 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
     5101 tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt
     5161 tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat cttattaagg
     5221 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5281 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5341 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5401 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5461 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5521 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5581 ctgcaacttt atccgcctcc atccagtcta ttccatggtg ccacctgacg tctaagaaac
     5641 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggcaga atttcagata
     5701 aaaaaaatcc ttagctttcg ctaaggatga tttctggaat tcgcggccgc ttctagagtg
     5761 gtgcaaaacc tttgcggtat ggcatgatag cg
//
