LOCUS       40924_34431        4619 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPh115, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4619)
  AUTHORS   Krom RJ, Bhargava P, Lobritz MA, Collins JJ.
  TITLE     Engineered Phagemids for Nonlytic, Targeted Antibacterial Therapies
  JOURNAL   Nano Lett. 15 (7), 4808-4813 (2015)
  PUBMED    26044909
REFERENCE   2  (bases 1 to 4619)
  AUTHORS   Krom RJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-JUN-2015) Molecular Medicine, Boston University, 72 
            East Concord St, Boston, MA 02118, USA
REFERENCE   3  (bases 1 to 4619)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4619)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nano 
            Lett."; date: "2015"; volume: "15"; issue: "7"; pages: "4808-4813"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-JUN-2015) Molecular Medicine, Boston University, 72 East Concord
            St, Boston, MA 02118, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        31..104
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     CDS             142..264
                     /codon_start=1
                     /product="cecropin PR-39"
                     /label=cecropin PR-39
                     /protein_id="AKQ98672.1"
                     /translation="MRRRPRPPYLPRPRPPPFFPPRLPPRIPPGFPPRFPPRFP"
     CDS             313..435
                     /codon_start=1
                     /product="cecropin PR-39"
                     /label=cecropin PR-39
                     /protein_id="AKQ98673.1"
                     /translation="MRRRPRPPYLPRPRPPPFFPPRLPPRIPPGFPPRFPPRFP"
     terminator      456..550
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      631..704
                     /label=PLtet0
                     /note="PLtet0"
                     /regulatory_class="promoter"
     promoter        631..704
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     protein_bind    631..649
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    656..674
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             730..738
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             745..804
                     /codon_start=1
                     /product="apidaecin"
                     /label=apidaecin
                     /protein_id="AKQ98674.1"
                     /translation="MGNNRPVYIPQPRPPHPRI"
     RBS             843..851
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             857..916
                     /codon_start=1
                     /product="apidaecin"
                     /label=apidaecin
                     /protein_id="AKQ98675.1"
                     /translation="MGNNRPVYIPQPRPPHPRI"
     regulatory      923..1045
                     /label=T0
                     /note="T0"
                     /regulatory_class="terminator"
     terminator      937..1031
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      1064..1137
                     /label=PLtet0
                     /note="PLtet0"
                     /regulatory_class="promoter"
     promoter        1064..1137
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1064..1082
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1089..1107
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1178..1480
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     regulatory      1490..1612
                     /label=T0
                     /note="T0"
                     /regulatory_class="terminator"
     terminator      1504..1598
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1619..1692
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1619..1637
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1644..1662
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1733..2035
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     terminator      2059..2153
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2186..2613
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     terminator      2638..2724
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      complement(2888..3476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3564..3658)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3692..4483)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 gacgtctgtg caagtactac tgttctgcag tccctatcag tgatagagat tgacatccct
       61 atcagtgata gagatactga gcacatcagc aggacgcact gaccggtacc ctataagtta
      121 cacaacataa ggaggaatat aatgcgtcgt cgtccgcgtc cgccgtacct gccgcgtccg
      181 cgtccgccgc cgttcttccc gccgcgtctg ccgccgcgta tcccgccggg tttcccgccg
      241 cgtttcccgc cgcgtttccc gtaaccgcgg agtatttgta cattctatca aagtatcata
      301 aggaggagaa taatgcgtcg tcgtccgcgt ccgccgtacc tgccgcgtcc gcgtccgccg
      361 ccgttcttcc cgccgcgtct gccgccgcgt atcccgccgg gtttcccgcc gcgtttcccg
      421 ccgcgtttcc cgtaaggatc ccgagaattg gcttggactc ctgttgatag atccagtaat
      481 gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat
      541 tggtgagaat ccaagcagta gtcagaattc gatacccagc tgggtggagt gcaccaagga
      601 gcatgcgaag gaaacgtttc gcagaagctt tccctatcag tgatagagat tgacatccct
      661 atcagtgata gagatactga gcacatcagc aggacgcact gaccggtacc caacagggat
      721 tcgttaattt aaggaggtct ccaaatgggc aacaaccgcc cggtgtatat tccgcagccg
      781 cgcccgccgc atccgcgcat ttaaccgcgg agtatttgta cagaggtaca actcaactaa
      841 tataaggagg ttcaaaatgg gcaacaaccg cccggtgtat attccgcagc cgcgcccgcc
      901 gcatccgcgc atttaaggat cccgagaatt ggcttggact cctgttgata gatccagtaa
      961 tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
     1021 ttggtgagaa tccaagcagt agtcagagct cggactgctt aagtccctat cagtgataga
     1081 gattgacatc cctatcagtg atagagatac tgagcacatc agcaggacgc actgaccggt
     1141 acctatccat cattaacgac caaatcaagg aggacgtatg cagtttaagg tttacaccta
     1201 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
     1261 cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg
     1321 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
     1381 ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
     1441 tgacatcaaa aacgccatta acctgatgtt ctggggaata taaggatccc gagaattggc
     1501 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
     1561 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcagtagt caggcgcctc
     1621 cctatcagtg atagagattg acatccctat cagtgataga gatactgagc acatcagcag
     1681 gacgcactga ccggtaccta tccatcatta acgaccaaat caaggaggac gtatgcagtt
     1741 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
     1801 tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
     1861 agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
     1921 gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct
     1981 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataagg
     2041 atcccgagaa ttggcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     2101 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagca
     2161 gtagtcacct agggaccgtc tcgagagcgc cctgtagcgg cgcattaagc gcggcgggtg
     2221 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2281 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2341 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2401 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2461 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2521 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2581 atgagctgat ttaacaaaaa tttaacgcga attaaatatt aatccaacgc gtggcatcaa
     2641 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     2701 aacgctctcc tgagtaggac aaatccgccg ccctagactt aggcgttcgg ctgcggcgag
     2761 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2821 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2881 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2941 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3001 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3061 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3121 ttcgctccaa gctgggctgt ctgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3181 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3241 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3301 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     3361 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3421 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3481 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3541 ttttggtcat ggctagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc
     3601 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca
     3661 agccaattct cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg
     3721 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     3781 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     3841 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     3901 tcggcaagca ggcatcgccg tgggtcacga cgagatcctc gccgtcgggc atacgcgcct
     3961 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     4021 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     4081 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     4141 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     4201 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagcc gcgcaaggaa
     4261 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
     4321 cggacaggtc ggtcttgaca aaaagaaccg ggcgaccctg cgctgacagc cggaacacgg
     4381 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     4441 aagccgccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
     4501 ctgtctcttg atcagatctt gatcccctgc gccatcagat ccttggcggc aagaaagcca
     4561 tccagtttac tttgcagggc ttcccaacct taccagaggg cggcccaact ggcaattcc
//
