LOCUS       sequence-224469-        6380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  surface-displayed Ag2, used in composability experiments; low copy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6380)
  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 6380)
  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: pDSG287
FEATURES             Location/Qualifiers
     source          1..6380
                     /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      2923..2994
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3010..3037
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3044..3064
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3065..3122
                     /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."
     primer_bind     complement(3146..3163)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(3425..4375)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4423..4645
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5137..5231)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5261..6052,6053..6121))
                     /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     5884..5903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6122..6226)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6291..6309
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(6313..6356)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    6359..6380
                     /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 cgactagaga accagaggct taataatact agagccaggc atcaaataaa acgaaaggct
     2941 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     3001 agtcacactg gctcaccttc gggtgggcct ttctgcgttt atatactagt agcggccgct
     3061 gcagtccggc aaaaaagggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa
     3121 gattacttcg cgttatgcag gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3181 tgcggcgagc ggtatcagct cactcaaagg cggtaatctc gaggttacat tgtcgatctg
     3241 ttcatggtga acagctttga atgcaccaaa aactcgtaaa agctctgatg tatctatctt
     3301 ttttacaccg ttttcatctg tgcatatgga cagttttccc tttgatatgt aacggtgaac
     3361 agttgttcta cttttgtttg ttagtcttga tgcttcactg atagatacaa gagccataag
     3421 aacctcagat ccttccgtat ttagccagta tgttctctag tgtggttcgt tgtttttgcg
     3481 tgagccatga gaacgaacca ttgagatcat acttactttg catgtcactc aaaaattttg
     3541 cctcaaaact ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc
     3601 cgttatgtag gtaggaatct gatgtaatgg ttgttggtat tttgtcacca ttcattttta
     3661 tctggttgtt ctcaagttcg gttacgagat ccatttgtct atctagttca acttggaaaa
     3721 tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa tttcatattg ctgtaagtgt
     3781 ttaaatcttt acttattggt ttcaaaaccc attggttaag ccttttaaac tcatggtagt
     3841 tattttcaag cattaacatg aacttaaatt catcaaggct aatctctata tttgccttgt
     3901 gagttttctt ttgtgttagt tcttttaata accactcata aatcctcata gagtatttgt
     3961 tttcaaaaga cttaacatgt tccagattat attttatgaa tttttttaac tggaaaagat
     4021 aaggcaatat ctcttcacta aaaactaatt ctaatttttc gcttgagaac ttggcatagt
     4081 ttgtccactg gaaaatctca aagcctttaa ccaaaggatt cctgatttcc acagttctcg
     4141 tcatcagctc tctggttgct ttagctaata caccataagc attttcccta ctgatgttca
     4201 tcatctgagc gtattggtta taagtgaacg ataccgtccg ttctttcctt gtagggtttt
     4261 caatcgtggg gttgagtagt gccacacagc ataaaattag cttggtttca tgctccgtta
     4321 agtcatagcg actaatcgct agttcatttg ctttgaaaac aactaattca gacatacatc
     4381 tcaattggtc taggtgattt taatcactat accaattgag atgggctagt caatgataat
     4441 tacatgtcct tttcctttga gttgtgggta tctgtaaatt ctgctagacc tttgctggaa
     4501 aacttgtaaa ttctgctaga ccctctgtaa attccgctag acctttgtgt gttttttttg
     4561 tttatattca agtggttata atttatagaa taaagaaaga ataaaaaaag ataaaaagaa
     4621 tagatcccag ccctgtgtat aactcactac tttagtcagt tccgcagtat tacaaaagga
     4681 tgtcgcaaac gctgtttgct cctctacaaa acagacctta aaaccctaaa ggcttaagta
     4741 gcaccctcgc aagctcgggc aaatcgctga atattccttt tgtctccgac catcaggcac
     4801 ctgagtcgct gtctttttcg tgacattcag ttcgctgcgc tcacggctct ggcagtgaat
     4861 gggggtaaat ggcactacag gcgcctttta tggattcatg caaggaaact acccataata
     4921 caagaaaagc ccgtcacggg cttctcaggg cgttttatgg cgggtctgct atgtggtgct
     4981 atctgacttt ttgctgttca gcagttcctg ccctctgatt ttccagtctg accacttcgg
     5041 attatcccgt gacaggtcat tcagactggc taatgcaccc agtaaggcag cggtatcatc
     5101 aacaggctta cccgtcttac tgtccctagt gcttggattc tcaccaataa aaaacgcccg
     5161 gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat
     5221 caacaggagt ccaagcgagc tcgtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5281 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5341 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5401 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5461 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5521 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5581 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5641 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5701 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5761 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5821 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5881 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5941 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6001 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6061 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6121 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6181 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttcccatgg
     6241 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6301 tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctgga
     6361 attcgcggcc gcttctagag
//
