LOCUS       sequence-455436-        3913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrates KanR into NS1, 500 bp homology, one homologous HIP1 site 
            in left homology arm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3913)
  AUTHORS   Kamoku C, Nielsen DR
  TITLE     Highly iterated palindrome 1 sequence improves Synechococcus sp. PCC
            7002 transformation efficiencies in a homology- and 
            methylation-dependent manner.
  JOURNAL   Biotechnol Prog. 2025 Jan 23:e3518. doi: 10.1002/btpr.3518.
  PUBMED    39846505
REFERENCE   2  (bases 1 to 3913)
  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: "Biotechnol 
            Prog. 2025 Jan 23:e3518. doi: 10.1002/btpr.3518."
COMMENT     Sequence Label: pCMK103
FEATURES             Location/Qualifiers
     source          1..3913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      176..764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     665..684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1532..2326
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1586..1605)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2196..2215
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      2357..2451
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 cgacgaccgg gtcgaatttg ctttcgaatt tctgccattc atccgcttat tatcacttat
       61 tcaggcgtag caaccaggcg tttaagggca ccaataactg ccttaaaaaa attacgcccc
      121 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
      181 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      241 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      301 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
      361 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
      421 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      481 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      541 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      601 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      661 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
      721 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
      781 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
      841 cccctgattc tgtggataac cgtattaccg cctttgagtg agcttcaagc ccaagggttt
      901 cgtagggatt ttgttcgctc attctcctcc tcctgtgagc atgccactgg gttattggaa
      961 catccttaga ttcttaatgt aacccaatca tcatacagtt tattattatg agcgttttcc
     1021 ggtgcgcttt ccatagagca aaattttttc tggtgatgat agccggggcg atcgcctcaa
     1081 gacttgccca gaccagagag tatacatccc catacttaac gctagctgat caacggagta
     1141 ggtactcgaa ccactcaggc tccctaaacc gctgcaatgt acaaaatctt agagaccatc
     1201 ccaaagaccc tccactaagc gatcgtgtag aggccactga acagacaagc acaaaaatac
     1261 aatcctccca gactatggtt tgtgtccgtt gatcgcactc ggcatgaaaa taagtccatt
     1321 ttgtcgattg accctaccat aaaaactcaa gcttgatctt ttatggatcc tctttctttc
     1381 gatctggcag acgtcggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
     1441 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcatgatc
     1501 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     1561 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     1621 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     1681 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     1741 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     1801 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     1861 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     1921 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     1981 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     2041 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     2101 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     2161 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     2221 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     2281 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     2341 ttcgagagct cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
     2401 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagcact
     2461 agtcattgct tcttttaacc tcagattaat taggcactaa ttttatagaa aaaagcttca
     2521 ttatatagaa tttctaaaaa tcaagcttta aagggttgac cttccttttg aaaggtaata
     2581 aaactgaagt acaaacaaat acgtaagtaa cgaaaaggag taagccgcaa tctgcctcag
     2641 tagaaacacg tatttttacc ccaaaagaag tattctattc atctttctcc tttgtgagaa
     2701 ggttcatgga gatccttctg gggcacttga aacctctact tgcaaggatc aaagcgacat
     2761 atgtacgtgt ttttccaaaa aaatcccaga agtagggagt aattcctcta ctgagatttc
     2821 aagaaaatta agtgttctga ggcttggttt atacgggaaa tttagacttg atggtttggg
     2881 tcttaccttt tcctagcagc aagactaaat taattagtta cgcatttgtt gaaatcaaac
     2941 tgaattctat tgaggattcc ttacaatggc taaattcaat tcttctacaa tcacattcgc
     3001 aatcgcacct cgctaacgga ttcaccactc caagaattgg agccaatcaa ttcttgcgga
     3061 gaactgtgaa tgcgcaaacc aacccttggc agaacatatc catcgcgtcc gccatctcca
     3121 gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg
     3181 tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg
     3241 aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag
     3301 caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtccc
     3361 ctacgtgctg ctgaagttgc ccgcaacaga gagtggaacc aaccggtgat accacgatac
     3421 tatgactgag agtcaacgcc atgagcggcc tcatttctta ttctgagtta caacagtccg
     3481 caccgctgcc ggtagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     3541 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgccca acagtccccc
     3601 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ccctgcacca ttatgttccg
     3661 gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa
     3721 gcgctaaccg tttttatcag gctctgggag gcagaataaa tgatcatatc gtcaattatt
     3781 acctccacgg ggagagcctg agcaaactgg cctcaggcat ttgagaagca cacggtcaca
     3841 ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga
     3901 ccctgccctg aac
//
