LOCUS       40924_15195        5826 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSG254, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5826)
  AUTHORS   Glass DS, Riedel-Kruse IH.
  TITLE     A synthetic bacterial cell-cell adhesion toolbox for programming 
            multicellular morphologies and patterns
  JOURNAL   Cell (2018) In press
REFERENCE   2  (bases 1 to 5826)
  AUTHORS   Glass DS.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-JUN-2018) Bioengineering, Stanford University, 318 
            Campus Drive, Clark Center E350, Stanford, CA 94305, USA
REFERENCE   3  (bases 1 to 5826)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5826)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell (2018)
            In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-JUN-2018) Bioengineering, Stanford University, 318 Campus Drive,
            Clark Center E350, Stanford, CA 94305, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..82
                     /label=lacI promoter
     CDS             83..1162
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    1233..1254
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1269..1299
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1307..1323
                     /label=lac operator
                     /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)."
     primer_bind     1331..1347
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1383..3350
                     /label=intimin N-terminus
                     /note="intimin N-terminus"
     misc_feature    3351..3389
                     /label=E-tag
                     /note="E-tag"
     CDS             3393..3407
                     /codon_start=1
                     /product="EPEA peptide protein"
                     /label=EPEA peptide protein
                     /note="C-terminus of alpha synuclein"
                     /protein_id="AXC07689.1"
                     /translation="EPEA"
     misc_feature    3393..3404
                     /label=Neae fusion
                     /note="Neae fusion"
     misc_feature    3420..3446
                     /label=myc tag
                     /note="myc tag"
     rep_origin      3545..4000
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             4187..4843
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     rep_origin      5178..5766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acccgacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
       61 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
      121 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
      181 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
      241 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
      301 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
      361 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
      421 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
      481 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
      541 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
      601 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
      661 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
      721 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
      781 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
      841 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gacatctcgg tagtgggata
      901 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
      961 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     1021 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
     1081 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     1141 ttcccgactg gaaagcgggc agtgagcggt acccgataaa agcggcttcc tgacaggagg
     1201 ccgttttgtt ttgcagccca cctcaacgca attaatgtga gttagctcac tcattaggca
     1261 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     1321 caatttcaca caggaaacag ctatgaccat gattacgaat ttctagataa cgagggcaaa
     1381 tcatgattac tcatggttgt tatacccgga cccggcacaa gcataagcta aaaaaaacat
     1441 tgattatgct tagtgctggt ttaggattgt ttttttatgt taatcagaat tcatttgcaa
     1501 atggtgaaaa ttattttaaa ttgggttcgg attcaaaact gttaactcat gatagctatc
     1561 agaatcgcct tttttatacg ttgaaaactg gtgaaactgt tgccgatctt tctaaatcgc
     1621 aagatattaa tttatcgacg atttggtcgt tgaataagca tttatacagt tctgaaagcg
     1681 aaatgatgaa ggccgcgcct ggtcagcaga tcattttgcc actcaaaaaa cttccctttg
     1741 aatacagtgc actaccactt ttaggttcgg cacctcttgt tgctgcaggt ggtgttgctg
     1801 gtcacacgaa taaactgact aaaatgtccc cggacgtgac caaaagcaac atgaccgatg
     1861 acaaggcatt aaattatgcg gcacaacagg cggcgagtct cggtagccag cttcagtcgc
     1921 gatctctgaa cggcgattac gcgaaagata ccgctcttgg tatcgctggt aaccaggctt
     1981 cgtcacagtt gcaggcctgg ttacaacatt atggaacggc agaggttaat ctgcagagtg
     2041 gtaataactt tgacggtagt tcactggact tcttattacc gttctatgat tccgaaaaaa
     2101 tgctggcatt tggtcaggtc ggagcgcgtt acattgactc ccgctttacg gcaaatttag
     2161 gtgcgggtca gcgttttttc cttcctgcaa acatgttggg ctataacgtc ttcattgatc
     2221 aggatttttc tggtgataat acccgtttag gtattggtgg cgaatactgg cgagactatt
     2281 tcaaaagtag cgttaacggc tatttccgca tgagcggctg gcatgagtca tacaataaga
     2341 aagactatga tgagcgccca gcaaatggct tcgatatccg ttttaatggc tatctaccgt
     2401 catatccggc attaggcgcc aagctgatat atgagcagta ttatggtgat aatgttgctt
     2461 tgtttaattc tgataagctg cagtcgaatc ctggtgcggc gaccgttggt gtaaactata
     2521 ctccgattcc tctggtgacg atggggatcg attaccgtca tggtacgggt aatgaaaatg
     2581 atctccttta ctcaatgcag ttccgttatc agtttgataa atcgtggtct cagcaaattg
     2641 aaccacagta tgttaacgag ttaagaacat tatcaggcag ccgttacgat ctggttcagc
     2701 gtaataacaa tattattctg gagtacaaga agcaggatat tctttctctg aatattccgc
     2761 atgatattaa tggtactgaa cacagtacgc agaagattca gttgatcgtt aagagcaaat
     2821 acggtctgga tcgtatcgtc tgggatgata gtgcattacg cagtcagggc ggtcagattc
     2881 agcatagcgg aagccaaagc gcacaagact accaggctat tttgcctgct tatgtgcaag
     2941 gtggcagcaa tatttataaa gtgacggctc gcgcctatga ccgtaatggc aatagctcta
     3001 acaatgtaca gcttactatt accgttctgt cgaatggtca agttgtcgac caggttgggg
     3061 taacggactt tacggcggat aagacttcgg ctaaagcgga taacgccgat accattactt
     3121 ataccgcgac ggtgaaaaag aatggggtag ctcaggctaa tgtccctgtt tcatttaata
     3181 ttgtttcagg aactgcaact cttggggcaa atagtgccaa aacggatgct aacggtaagg
     3241 caaccgtaac gttgaagtcg agtacgccag gacaggtcgt cgtgtctgct aaaaccgcgg
     3301 agatgacttc agcacttaat gccagtgcgg ttatattttt tgatggtgcg ccggtgccgt
     3361 atccggatcc gctggaaccg gcccagccgg ccgaaccaga ggcttaataa gcggccgccg
     3421 aacaaaaact catctcagaa gaggatgcag ctgcataaaa gcttgacctg tgaagtgaaa
     3481 aatggcgcac attgtgcgac attttttttg tctgccgttt accgctactg cgtcacggat
     3541 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3601 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3661 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     3721 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3781 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3841 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3901 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3961 tttaacgcga attttaacaa aatattaacg tttacaattt caggtggcac ttttcgggga
     4021 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4081 atgtcgagac gttgggtgag gttccaactt tcaccataat gaaataagat cactaccggg
     4141 cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     4201 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     4261 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
     4321 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     4381 cctgatgaat gctcatccgg agttccgtat ggcaatgaaa gacggtgagc tggtgatatg
     4441 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     4501 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     4561 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     4621 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     4681 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
     4741 gccgctggcg attcaggttc atcatgccgt ctgtgatggc ttccatgtcg gcagaatgct
     4801 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag
     4861 ttattggtgc ccttaaacgc ctggtgctac gcctgaataa gtgataataa gcggatgaat
     4921 ggcagaaatt cgaaagcaaa ttcgacccgg tcgtcggttc agggcagggt cgttaaatag
     4981 ccgcttatgt ctattgctgg tttaccggtt tattgactac cggaagcagt gtgaccgtgt
     5041 gcttctcaaa tgcctgaggc cagtttgctc aggctctccc cgtggaggta ataattgctc
     5101 gacatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5161 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5221 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5281 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     5341 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5401 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5461 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5521 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5581 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5641 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5701 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5761 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5821 cacatg
//
