LOCUS       sequence-224791-        6746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  surface-displayed Nb2, used in composability experiments; low copy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6746)
  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 6746)
  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: pDSG289
FEATURES             Location/Qualifiers
     source          1..6746
                     /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."
     primer_bind     complement(3512..3529)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(3791..4741)
                     /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      4789..5011
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5503..5597)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5627..6418,6419..6487))
                     /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     6250..6269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6488..6592)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6657..6675
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(6679..6722)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    6725..6746
                     /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 aatctcgagg ttacattgtc
     3601 gatctgttca tggtgaacag ctttgaatgc accaaaaact cgtaaaagct ctgatgtatc
     3661 tatctttttt acaccgtttt catctgtgca tatggacagt tttccctttg atatgtaacg
     3721 gtgaacagtt gttctacttt tgtttgttag tcttgatgct tcactgatag atacaagagc
     3781 cataagaacc tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt
     3841 tttgcgtgag ccatgagaac gaaccattga gatcatactt actttgcatg tcactcaaaa
     3901 attttgcctc aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc
     3961 ttagtccgtt atgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca
     4021 tttttatctg gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt
     4081 ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt
     4141 aagtgtttaa atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat
     4201 ggtagttatt ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg
     4261 ccttgtgagt tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt
     4321 atttgttttc aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga
     4381 aaagataagg caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg
     4441 catagtttgt ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag
     4501 ttctcgtcat cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga
     4561 tgttcatcat ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag
     4621 ggttttcaat cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct
     4681 ccgttaagtc atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca
     4741 tacatctcaa ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat
     4801 gataattaca tgtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg
     4861 ctggaaaact tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt
     4921 tttttgttta tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa
     4981 aaagaataga tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca
     5041 aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct
     5101 taagtagcac cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc
     5161 aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca
     5221 gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc
     5281 ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt
     5341 ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca
     5401 cttcggatta tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt
     5461 atcatcaaca ggcttacccg tcttactgtc cctagtgctt ggattctcac caataaaaaa
     5521 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     5581 atctatcaac aggagtccaa gcgagctcgt aaacttggtc tgacagttac caatgcttaa
     5641 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5701 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5761 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5821 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5881 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5941 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6001 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6061 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6121 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6181 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6241 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6301 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6361 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6421 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6481 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6541 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6601 ccatggtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     6661 ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt agctttcgct aaggatgatt
     6721 tctggaattc gcggccgctt ctagag
//
