LOCUS       40924_20476        4798 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Integrative expression vector pFPN, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4798)
  AUTHORS   Chaurasia AK, Parasnis A, Apte SK.
  TITLE     An integrative expression vector for strain improvement and 
            environmental applications of the nitrogen fixing cyanobacterium, 
            Anabaena sp. strain PCC7120
  JOURNAL   J. Microbiol. Methods 73 (2), 133-141 (2008)
  PUBMED    18367274
REFERENCE   2  (bases 1 to 4798)
  AUTHORS   Chaurasia AK, Parasnis A, Apte SK.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-MAR-2007) Molecular Biology Division, Bhabha Atomic 
            Research Centre, Trombay, Mumbai, Maharashtra 400 085, India
REFERENCE   3  (bases 1 to 4798)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4798)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Microbiol. Methods"; date: "2008"; volume: "73"; issue: "2"; pages: 
            "133-141"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (05-MAR-2007) Molecular Biology Division, Bhabha Atomic Research 
            Centre, Trombay, Mumbai, Maharashtra 400 085, India"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    707..2564
                     /label=integrative expression cassette
                     /note="integrative expression cassette"
     misc_feature    721..1221
                     /note="F1 flanking region; derived from Anabaena sp.
                     PCC7120"
     regulatory      1224..1388
                     /note="light inducible PpsbA1 promoter from Anabaena sp. 
                     PCC7120"
                     /regulatory_class="promoter"
     CDS             1458..2189
                     /codon_start=1
                     /gene="nptII"
                     /product="neomycin phosphotransferase"
                     /label=nptII
                     /note="derived from pDUCA7"
                     /protein_id="ABO72539.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRPPILKPITILLGSEDLP"
     gene            1458..2189
                     /gene="nptII"
                     /label=nptII
     misc_feature    2139..2570
                     /note="F2 flanking region; derived from Anabaena sp.
                     PCC7120"
     promoter        complement(2607..2625)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2646..2662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2670..2686)
                     /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)."
     promoter        complement(2694..2724)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2739..2760)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3048..3636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3810..4667)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4668..4772)
                     /label=AmpR promoter
ORIGIN
        1 ggaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg cgccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcgact tcacgccata
      721 atcatgtgtc aaaaattcct tatatattaa tgaattttat cttataaaac aatattattg
      781 tttactctac gcttcacata gaaattgtga aaagctcttt tccagacgct tagaacttag
      841 ggacaaaaat gatttctgga tacttgcgct caccgtgtag tacgtttcca gtggcaattg
      901 ttcagtctgt agcagatatt caaactgatt gactttacga ctgaacagct tgataggctt
      961 agatagtatt gctattaagt ttcagtttgg tcacaaaaag taacaaatta attaatcagt
     1021 cagattttta ttcctttgtt atctttaaat tttcgcatca ataaataaca cagataggag
     1081 ttgactttga tttctgttgg cgcagtgtgg catagctata ggtacttgtg tatctagggg
     1141 atggtcactc cgaaccaaca aaatttaaat gggatgccta gatgaagtct aattgtatgg
     1201 ttagagttgt aattatcccc actgcaggga ttcccaaaga tagggggaat aattaacatt
     1261 aagaattatt aattcatggg tttttagtct agtaaatttg cgtgaattca tgtaaatttt
     1321 atgagacagg cgcaagtcta aaaaaagcgt ctgaattaat ctgcacaaat ccaaagcaat
     1381 cataaaaaca tatggggatc cgcggggatc aagatccccg gggatctgat caagagacag
     1441 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     1501 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     1561 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     1621 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     1681 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     1741 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     1801 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     1861 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     1921 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     1981 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     2041 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     2101 cggaccgcta tcaggacata gcgttggcta cccggccccc aattttgaaa cctatcacta
     2161 tattattagg cagtgaagac ttgccataac ctatatttct cgcatgaagt aatctgctga
     2221 gaactatgag ttgtgtaata tattgtattt ttattaagtg taggtttaat tgcatagagt
     2281 tagccgttag aactaattat cccagaatag atacctcaaa ttctcagtag cagaaattaa
     2341 atagataaca aaatatacaa aatgccttca gtatttaaaa cttcttctca gaagacaaac
     2401 acagttagta accagatgaa taactcatca aaatcatcaa cagaagtatt agctgtgtct
     2461 cctgcatccc aaaggtatac tatgcaaaaa ttgccatgta gagacaatat tagtaggttg
     2521 tggaagttat acagatcaat tttaagtaaa tgtatgtata cttctctaga gcggccgcca
     2581 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattccgag cttggcgtaa
     2641 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2701 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2761 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2821 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2881 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2941 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3001 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3061 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3241 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3481 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3541 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3661 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3721 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3781 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3841 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3901 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3961 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4021 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4081 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4141 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4201 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4261 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4321 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4381 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4441 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4501 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4561 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4621 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4681 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4741 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg
//
