LOCUS       sequence-234228-        4372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cyanobacteria cloning vector. Chromosomal integration into 
            Synechococcus elongatus neutral site 2 (NS2)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4372)
  AUTHORS   Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B, 
            Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW
  TITLE     Broad-host-range vector system for synthetic biology and 
            biotechnology in cyanobacteria.
  JOURNAL   Nucleic Acids Res. 2014 Jul 29. pii: gku673.
  PUBMED    25074377
REFERENCE   2  (bases 1 to 4372)
  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: "Nucleic 
            Acids Res. 2014 Jul 29. pii: gku673."
COMMENT     Sequence Label: pAM4937 (pCV0049)
FEATURES             Location/Qualifiers
     source          1..4372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      504..1092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     993..1012
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1246..1263
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1278..1418
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1344..1363)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(2552..3367)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2552..3367)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3275..3294
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3359..3378
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3937..4242
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4121..4140
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     terminator      4310..4337
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tctagagttg ctaccaagta gtgacccgct tcgtgatgca aaatccgctg acgatattcg
       61 ggcgatcgct gctgaatgcc atcgagcagt aacgtggcac cccgcccctg ccaagtcacc
      121 gcatccagac tgaacagcac caagaggcta aaacccaatc ccgccggtag cagcggagaa
      181 ctacccagca ttggtcccac caaagctaat gccgtcgtgg taaaaatcgc gatcgccgtc
      241 agactcaagc ccagttcgct catgcttcct catctaggtc acagtcttcg gcgatcgcat
      301 cgatctgatg ctgcagcaag cgttttccat accggcgatc gcgccgtcgc cctttcgctg
      361 ccgtggcccg cttacgagct cgtttatcga ccacgatcgc atccaaatcc gcgatcgctt
      421 cccagtccgg caattcagtc tggggcgtcc gtttcattaa tcctgatagc gtcagacccc
      481 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
      541 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
      601 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
      661 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
      721 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
      781 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
      841 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
      901 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      961 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1021 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     1081 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     1141 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     1201 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     1261 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     1321 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     1381 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     1441 ctgacgcgcc ctgacgggct tgtctgctcc cgccatccgc ttacagacaa gctccgcggg
     1501 ccacgatttg agggacgaat cttcagcgtg actgaagtac ctcgcggcaa gccatttccc
     1561 gatgtctttt tgttggccgc cgatcgcttc ggggttaatc ctacggcctg cgctgtgatc
     1621 gaagacaccc ccttgggagt agcggcaggc gtggcggcag gaatgcaagt gtttggctac
     1681 gcgggttcca tgcccgcttg gcgtctgcaa gaagccggtg cccatctcat ttttgacgat
     1741 atgcgactgc tgcccagtct gctccaatcg tcgccaaaag ataactccac agcattgccc
     1801 aatccctaac ccctgctcgc gccgcaacta cacactaaac cgttcctgcg cgatcgctct
     1861 tactgttgat ggctcgtgct taaaaacaat gcaaccctaa ccgtttcagc tggtgatttt
     1921 cggacgattt ggcttacagg gataactgag agtcaacagc ctctgtccgt cattgcacac
     1981 ccatccatgc actggggact tgactcatgc tgaatcacat ttcccttgtc cattgggcga
     2041 gaggggaggg gaatcttctg gactcttcac taagcggcga tcgcaggttc ttctacccaa
     2101 gcagtggcga tcgcttgatt gcagtcttca atgctggcct ctgcagccat cgccgccacc
     2161 aaagcatcgt aggcgggacg ttgttgctcc agtaaagtct tcgcccgtaa caatccccag
     2221 cgactgcgta aatccgcttc ggcaggattg cgatcgagtt gccgccacag ttgtttccac
     2281 tgggcgcgat cgtcagctcc cccttccacg ttgccgtaga ccagttgctc tgccgctgca
     2341 ccggccatca acacctgaca ccactgttcc agcgatcgct gactgagttg cccctgtgcg
     2401 gcttcggctt ctagcgcagc tgcttggaac tgcacacccc cgctagcatc gcgcgcgcgc
     2461 gcgcggatgc tagccaatgg cttattttcc tgctaattgg taataccatg gtaccgctct
     2521 gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa
     2581 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     2641 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     2701 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     2761 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
     2821 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     2881 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
     2941 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     3001 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     3061 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     3121 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     3181 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     3241 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     3301 aatcagcatc catgttggaa tttaatcgcg gcctggagca agacgtttcc cgttgaatat
     3361 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
     3421 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggaccggta
     3481 tcccgccgcc gccgccggat accggttaat agtacctgtg aagtgaaaaa tggcgcacat
     3541 tgtgcgacat ttttttctcg agcaggtagt cggccaataa cccagggatt taaatacgcg
     3601 tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg
     3661 tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag
     3721 cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg
     3781 tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg
     3841 aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga
     3901 acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat
     3961 aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc
     4021 gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt
     4081 gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg
     4141 gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat
     4201 gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata
     4261 cacagccagt ctgcaggtcg acatttaaat gaaggagctc cccggcagga gaaggccatc
     4321 ctgacggatg gcctttttga agctttctag aatcgggggc ccccgggggg at
//
