LOCUS       sequence-457912-        9835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector suitable for transfer to Anabaena (Nostoc) sp. strain
            PCC 7120 and other cyanobacterial strains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9835)
  AUTHORS   Wolk CP, Cai Y, Cardemil L, Flores E, Hohn B, Murry M, Schmetterer 
            G, Schrautemeier B, Wilson R
  TITLE     Isolation and complementation of mutants of Anabaena sp. strain PCC 
            7120 unable to grow aerobically on dinitrogen.
  JOURNAL   J Bacteriol. 1988 Mar;170(3):1239-44.
  PUBMED    2830231
REFERENCE   2  (bases 1 to 9835)
  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"; journalName: "J 
            Bacteriol."; date: "1988-03"; volume: "170(3)"; pages: "1239-44"
COMMENT     Sequence Label: pRL25
FEATURES             Location/Qualifiers
     source          1..9835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             739..1533
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(793..812)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1403..1422
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1733..1752)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1972..1994)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2080..2220
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2135..2154
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2235..2252)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2406..2994)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2486..2505)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        complement(join(3406..3411,3412..3428,3429..3434))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 atcgtccatt ccgacagcat cgccagtcac tatggcgtgc tgctagcgct atatgcgttg
       61 atgcaatttc tatgcgcagt gggcgcatca cgctaaaaca ggggaatatc acgctcaacg
      121 cggtgctggc cgaggagatt aacccgccca agggtgagac cccgttgaaa tggttgttgc
      181 tgaccagcga accggtcgag tcgctagccc aagccttgcg cgtcatcgac atttataccc
      241 atcgctggcg gatcgaggag ttccataagg catggaaaac cggagcagga gccgagaggc
      301 aacgcatgga ggagccggat aatctggagc ggatggtctc gatcctctcg tttgttgcgg
      361 tcaggctgtt acagctcaga gaaagcttca cgctgccgca agcactcagg gcgcaagggc
      421 tgctaaagga agcggaacac gtagaaagcc agtccgcaga aacggtgctg accccggatg
      481 aatgtcagct actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta
      541 gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa
      601 ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg
      661 atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga tcaagagaca
      721 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
      781 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
      841 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
      901 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
      961 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     1021 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     1081 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     1141 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     1201 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     1261 aaggcgcgca tgcccgacgg cgatgatctc gtcgtgaccc atggcgatgc ctgcttgccg
     1321 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     1381 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     1441 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     1501 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     1561 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     1621 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     1681 tcatgctgga gttcttcggg catcaccggc gccacaggtg cggttgctgg cgcctatatc
     1741 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
     1801 ggcgtgggta tggtggcagg cccgctttat cagaagccag acattaacgc ttctggagaa
     1861 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
     1921 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     1981 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2041 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     2101 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     2161 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     2221 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2281 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2341 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2401 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2461 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2521 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2581 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2641 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2701 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2761 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2821 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     2881 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2941 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3001 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3061 ttggtcatga gattatcaaa aaggatcctt ttaaattaaa aatgaagttt taaatcaatc
     3121 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3181 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3241 actacgatac gggagggctt accatctggc ccggcagtac cggcataacc aagcctatgc
     3301 ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac
     3361 acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa agcttatcga
     3421 tgataagctg tcaaacatga gaattcttga agacgaaagg gcctcttgcg ggatatccgt
     3481 ctagatagtc atcgctcatc cctcacctca accgttttgc tgcctaacga ggtagtctaa
     3541 gaccgcgcca attctgtcga gtgccgcgct agtggtacgt tggtactcat caagccgtgc
     3601 ttcggtttgt aattgcctag cggtaagaat ttcaaagttc tgctgatgga tttgaactgt
     3661 ttgcaacaat gcttctgcgg ttgcgcggag ttctgtaacc gatgtgcgga ggtcggcatt
     3721 cactgcaact tggctgtcta acgcagcttc tattctggct agtctgtctt cgctcatgct
     3781 gccccctgct tgtcttggtt gtccttggct tcttgttcca tttgttcttt aacaactcgt
     3841 gtgagtaaga aagcggcaag attagccaac ggtcttcctt cctcagacgc tcggttagca
     3901 atccattggt aaatttcagc ttctagagtg actgctattc tccctttatt ggttggcatg
     3961 acgtttggtg tactcctgag tccatatcat acaccttgtg caaagagtga ttgacaaatg
     4021 tactaatcgg cagtaatagt atagagtgta ttaagtgtac accgagtata tgaccgaagt
     4081 acagaccgtc aacaatggca ctgtacccgt ttttaatgcg ggtaaaagtg ccagtaaaaa
     4141 actgcgtcgt aatctgcctt cgctgccatc caacccaggc gcgagtgaga tgagtgacgt
     4201 gagctattcc tgcaaagttg atgccagccc gactgtgtgg gatgctttac cgctatacag
     4261 cccgtttagc ttatcggctg atggcagtta cccaatggtc aaggtgacgc gctcaaaagc
     4321 ctgcgaactg agaaccggga aatcaatcct ggtcggttcg ggtcgttgct accgagtgat
     4381 tttttgatga cgattcacct taaaggtgaa tcgcactgat tcctgtggac tgcctcgtaa
     4441 acacccactg gaatcgccta cataatgaag gacataatta tcatgacaga tagaccactc
     4501 accgagaaag agcagcaagt catctctgag tttgaaacgg cgcgtcctgg cttgggtgca
     4561 attgccgaac aaaacattat taataataat gtcaccggct gggctgacat tatcgctgat
     4621 actccggctg aagaactcgt cgccaatgaa gggactgcca gcaacagttt tatgtaccgg
     4681 agaatcggct aatgaacgag cgactttacc aagtatcctt gacagagtta actaaagttg
     4741 gtgtcccgca agcgatagcc gaacaagcgt ccttagtcgt agccaacgat gaccctacaa
     4801 aacccaatct tggcagaaca ccccaagccc aagaagcggt caatgcggct atgactcact
     4861 accacgctaa tcaacaggga gtaagtcaat aatgagcgat cgccagtttc tatttttagg
     4921 actggcggcc gccgccgtca gtctgtggta tcaaaaccag caaacagagt tacaagcaca
     4981 gttaaatacc tgtcgcgctc aactgcaagg ctttaaagac ggggtgattt atggcaaatg
     5041 actggattga aatgcacttg cctagtcgtc aacagctagg caggttttta ctgtggggat
     5101 tggggcgatt aacttttagt gccagcatat ttttatttga tgccacgtct tacgctttgc
     5161 aaacagcgtt aacagtggtg aagtggtgta gacagtctgt atacaacttg gtcaaagttt
     5221 acgaccaatt gccctatgct ggcacttacg gcgcggctat cattgacact accgccgtca
     5281 cccttgcggt cgaaccagag ggaatcatca ccgacttgat gcaagctctg tctggtaagc
     5341 actgtttaat agttggggat acgggaactg gcaaaagcac catagcccaa tggcttgctt
     5401 atcaagtcgg gggtgaggtg acagtctatg acgctgacgc tgcaccggac gagtggacgg
     5461 ggttaaatgt cattggccgc cgtggtgact ttgaggctat acagtcgggc atggctgctg
     5521 acctggagga attgcagcgc cggattgagc tacggggtga gtctggggat aaggcactag
     5581 caggaaaaga aagcgtttta attgctgaag aactgccact gctgagagat gaagtcgaga
     5641 tagcaaccga atggctcatc aaacacgcca ggcgcggccg caagcccaag cgttttgtta
     5701 ttgggctgac tcaagatgat aatgtcaaga ccttaggtat cgagggtgaa ggcggtgtcc
     5761 gtaagtgttt taggatgtta cgcctgggta agtttgcagt tagccacgcc aagtccttaa
     5821 aagaccttgc ccttgtggag tggttaaagt cggggaaata tcgctgcatg gtggatgata
     5881 tgccgtgtca gttaccagat gtaagcagtt acaaaatggt tatgccccgg ttacagtctg
     5941 taaccagttt tcagccactc caaacggctc aaccgacctc agaacaagca ttacagcccg
     6001 tttcatccct cccagaacca agagatgaaa ccctagcaaa agtggtaaaa gcctgtctgg
     6061 aagcgggttt ttcagaaagt aaggtgatta aagaaatttt gggctatcaa ggcgggcgtt
     6121 accaagaagg taaatcgcta ctggaaaaac ttaaggattt gtagcggtgc ggtttagcat
     6181 tgactgttac ttgctgtaca aggataattt tctcaaagcg gaatggtgtt aaacccgatt
     6241 attagagtcg tcatagcaaa agcttgaaaa ctggcaaggg tgtactggtt ttcgttcttt
     6301 ttggttctaa tggttatttt ttaccagttt atttagtaat aaccgagtgc caagtttctt
     6361 gttttacata gcaaagttag caaaaaacat tttgccgtga atttccatct gtaaaaagcg
     6421 tcacggcgaa attcagtatt tagtaaatac aaaaataagt caaacgttga tcatgcaaca
     6481 ctttggcgta caattgattg aaaattttgc cgataggcaa agtttaaatt acctattaaa
     6541 aaaagttcag tgcttggggg ttgttatgct acttaaccga agctaaataa aacacacata
     6601 cacagaattg ttaccacacc aaacaccaaa cagaaaaaac gtgctttgaa tttatcaatt
     6661 aatgggtgta gttaacaagt tttatttagc ttcaggttaa gcaaataaca ttcaataggt
     6721 actggactga gacatttcca aatgatcagc ccagaaagct taaatacaaa tgtatcacta
     6781 accctgctta caagggaact acaaactgaa ataattcttg gcacttacac ggtcagggta
     6841 cacactcgca ttggtcgaga accttgtgcg cggttgtggt atttgtgtcg agcgcttgat
     6901 aaggatgggt ctggtcattt aacattgcca ttgcctgtag tacagacgtt cttagattgt
     6961 agtgataagt ctgtttaccg ttggttacag gatggaaaaa aaataggagc attccgccgc
     7021 tacaaaatta aagcaggaat gataaccgtc tacttgggcg gtatgtttca agtttgttac
     7081 aacctaaatt taaaaagatg gggtgatgtt gcagtagtgc ctttagtcca agtgttaagc
     7141 gatttacgct cactgaccac tggcatagtc acacaaagct ttcaacaaaa gagccgctat
     7201 gcggccaaca gacaacttaa accagagtac agaaaacttt ttggagcgcc acacccgaac
     7261 gagctagtga aggacacaag gcaatcctct cttaaatcgc cagaggggga agtaccttgt
     7321 gtacttcaca ttagttcgtc gcggatattc gtctcaaaaa gtttcatcca ctatggtaca
     7381 tctcaaaaag ctgtcagctg tgagttgggt attcacaagc gcacagtgcg acgacatcaa
     7441 aagcagctgg gcatgaatcg ccgtcaactt tgccaagcca aaattgagta caatcaactc
     7501 cggcacgctc ggaataatga cgcgtctgaa ttttgggctt tcactggaac caaaactgat
     7561 atcggttatc aagttatggg tgacgcggtt gtgttttctg atggcattgc ccctggagcc
     7621 aaaaagagac agcccaacac ttaccaaatt gacgcgactg aatttgacgg tcgattattc
     7681 aaagtgggtg acaaggtgtt catgaatcgg tgcaacatct accgcgaaca attcacccta
     7741 acaacgatgt cagcagcacg gcggaagtac catttcaaac tgtcacagtg ccacttcagc
     7801 gaaaatcgag ccggtcgggt aggcaaccgt tttgtaatag gttgccactc tggcgagata
     7861 tgaaggacat ttttaagaat ttttttttag cgtgaaatgc tgcggcaacc agtatagtga
     7921 tcgacacgaa ctagtgcagc atgaccgaga gcctataggg tcgcggcttt ttggcaatag
     7981 agatcgtatg cagatcatgc gtagtacttt cttcgacctt aaccccaaag aaagcttatt
     8041 agatggagaa ggtcacaatt agaaaatatt catcgtcccg attgaccctt aacctttaat
     8101 cggtttccac ttgtaaaaaa ttcggcgagc cattcttctt aaccctcaga gtcgaacttc
     8161 atacaagaat atacaaaccc tttgggttta ccactaaaac aacggcgata aaaacctaat
     8221 aggtttccac agaaacgtct aaggatttag gcgtgttgtt ttggggcaat ttgagaaaat
     8281 attaaaccag tcacaagtta acttgtcgaa gaaatattgc agcattctgc aacacaggga
     8341 aaggtataac gtttccaaag tccggaacgt gtgcagagtt ctagaaccga tggatacttt
     8401 aaaagtatcc tcaagtgtag agcgtgtgtt taggctgcgt ctttttttag cacccatagg
     8461 ggtctaaaca gcgttggcgt acattagaag ccaatcacgg atacagcttt gcgtcgctgt
     8521 tcgggtgtaa cttcaagata gcgttgcagt gtgccaaggt cattgtgacc ggatatctct
     8581 tgaatgtgtc gcaacgggat accggcgcta gacatttggt tgagggcagt acggcggaaa
     8641 ctgtgggtac tgatgccttc tagcccgatt cttttggctg catcgcgcaa gattttatcc
     8701 gccgcgtatc gcgtgagcct atcgctgact cccctcatgc cagggaacag ggttcccggt
     8761 ttggggtgat agtcagccat gagtgcggct agtcctggct ggatgtcaac cacgcgggtt
     8821 ttgagtttcc ctttggtggt agacttccta aaggttagtg tttcgccttt aatgtccgtt
     8881 gtttggagtg ctagagcttc actaacgcgg caaccggtga atagacagat gccaaacaaa
     8941 acgcgatcgc gcggtgtggt gaatccgtcg ctaaacagtc gcctgagttc gtcggaggtg
     9001 agtattttgg ctcgtccgtt gccgttaacc ttcataaccc ttacagatta agcattccaa
     9061 tactctacac gttatctgca ttttgtggaa ttggtggcta tatctattga ttcttcgtat
     9121 gctttctgtt gaaagccagt gctagtttag ccaagcttat caatcgttgc acggtaaaat
     9181 caggctttcg acgtaaggac agaattttga taacttcagc ttatgttgtc tcctgaagaa
     9241 agcaactctt tgataaacgc gtcgatggct ttcccggcgg ctgactgcct acccattttt
     9301 agcttattga gggtgcgatc gcgggttgcc tccaatttgt ctagattgtc tatactattg
     9361 tctatattct ttctgtcttg cccggtagct tgtccttctg cctgaactac cgggctttcg
     9421 attttccccg cagttcctcc agttgcgatt gaagtttgtc tagacgtgtt gctagattat
     9481 cttctatacg cttgtctata aattcatcta tattgtctag attattagtt gtgactttat
     9541 ctagacgttg ttcaatattg tctatacgct gttcgaggga tgtgtctaga tgggtgggag
     9601 tgtgagaagt gggagtatta aacctgttac ctgccaaata ccaacgaaca aattctgtta
     9661 acagtgctga ggcgttggaa ctttcagaac tagcaaggtt tttaaaagac tcccattctt
     9721 cggagtcaat acgaaaagtg gctagagttt tgtctgccat gattagataa tgtctatact
     9781 tgtatagata gcagtataaa ctattgtcta tattagtgtc tatataagtc tagat
//
