LOCUS       40924_34426        4447 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPh113, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4447)
  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 4447)
  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 4447)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4447)
  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..4447
                     /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="AKQ98666.1"
                     /translation="MRRRPRPPYLPRPRPPPFFPPRLPPRIPPGFPPRFPPRFP"
     CDS             313..435
                     /codon_start=1
                     /product="cecropin PR-39"
                     /label=cecropin PR-39
                     /protein_id="AKQ98667.1"
                     /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="AKQ98668.1"
                     /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="AKQ98669.1"
                     /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"
     CDS             1525..1827
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     regulatory      1837..1959
                     /label=T0
                     /note="T0"
                     /regulatory_class="terminator"
     terminator      1851..1945
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2014..2441
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     terminator      2466..2552
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      complement(2716..3304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3392..3486)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3520..4311)
                     /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 taaccgcggt accaaccccc
     1501 ctattagaat aaggagtatc acgcatgcag tttaaggttt acacctataa aagagagagc
     1561 cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg
     1621 gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg
     1681 gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg
     1741 gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac
     1801 gccattaacc tgatgttctg gggaatataa ggatcccgag aattggcttg gactcctgtt
     1861 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
     1921 ccgggcgttt tttattggtg agaatccaag cagtagtcac aattgtcccc cggcgccagg
     1981 gttgatatct atcgccctag ggaccgtctc gagagcgccc tgtagcggcg cattaagcgc
     2041 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2101 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2161 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2221 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2281 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2341 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2401 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taaatattaa tccaacgcgt
     2461 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     2521 tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc ctagacttag gcgttcggct
     2581 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2641 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2701 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2761 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2821 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2881 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2941 gtaggtcgtt cgctccaagc tgggctgtct gcacgaaccc cccgttcagc ccgaccgctg
     3001 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3061 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3121 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     3181 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3241 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3301 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3361 ttaagggatt ttggtcatgg ctagtgcttg gattctcacc aataaaaaac gcccggcggc
     3421 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     3481 ggagtccaag ccaattctcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     3541 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     3601 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     3661 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     3721 catgatattc ggcaagcagg catcgccgtg ggtcacgacg agatcctcgc cgtcgggcat
     3781 acgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     3841 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     3901 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     3961 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     4021 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagccgc
     4081 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt
     4141 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgaccctgcg ctgacagccg
     4201 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     4261 ctccacccaa gccgccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     4321 tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa
     4381 gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg gcccaactgg
     4441 caattcc
//
