LOCUS       sequence-455437-        3880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrates KanR into NS2, 500 bp homology, one homologous HIP1 site 
            in right homology arm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3880)
  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 3880)
  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: pCMK104
FEATURES             Location/Qualifiers
     source          1..3880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      88..676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     577..596
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1445..2239
                     /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(1499..1518)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2109..2128
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      2270..2364
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 caccaataac tgccttaaaa aaattacgcc ccgtgagttt tcgttccact gagcgtcaga
       61 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      121 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
      181 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
      241 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
      301 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
      361 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
      421 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
      481 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
      541 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
      601 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      661 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      721 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      781 cgcctttgag tgagcttatt gggggcttgt tgaagattta ctttaggcat ttacgcttta
      841 actccttaaa tctaaagaac gaagaactat atggtgcggc ccaggggggc agatacttta
      901 caaaacaatc atcatgtagg tcacaatcta gaacacaaag aatttttcta aaaaataaac
      961 gacataccct ttaactttct tacgctttat aacttttaat tgagcgctca ctactttaga
     1021 cgggcaacaa tcgaaaaagg ttcaacgatt cattaaaatc tcagggatgg ggaattcatc
     1081 ggcgttctat acttaattct ccttgattga ccaagagctt gaaaagggga aaagatgcgg
     1141 gattaccgaa cttggaatat cgacagactc aattagaaat atagtgtcgt tgcaatatgt
     1201 tattcaatgc gataagggtg gcccttgcgg ggaagacatc tctcaaaaat gagacctcta
     1261 ttgcagtaga ggtacaggga gatgcccata aattaagact tagacgtcgg aattgccagc
     1321 tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
     1381 gccaaggatc tgatggcgca ggggatcatg atctgatcaa gagacaggat gaggatcgtt
     1441 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     1501 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     1561 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     1621 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     1681 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     1741 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     1801 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     1861 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     1921 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     1981 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     2041 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     2101 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     2161 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     2221 tcttgacgag ttcttctgag cgggactctg gggttcgaga gctcgcttgg actcctgttg
     2281 atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc
     2341 cgggcgtttt ttattggtga gaatccaagc actagtcatt ccttgtgatg atcactaccg
     2401 tttaaaagat tgaaaaagaa tatttttttg accaataagt ccagtttatt caaaaaaagg
     2461 cgctttgtat acctttagga aaaagataat ctcagcaaaa ataagggcat ttaggccaga
     2521 aatgaagaac gtagaagtat tttggcaagc cagggatttc acctaaaaat taacggttga
     2581 taatccttct caaaaataaa agaatgtcct tgttagtcca ggaaaccgag gtggcgatcg
     2641 cctttggatc aagatctatt cccagagttg cagaatttgt gccgtgatgg gatggcgttt
     2701 cgatttttgt aacccggtgc gatggattgg tcaggtgagt agcgatggat aatttttccg
     2761 gcctcttcac cccaaaaaga tctagcggct tccctggggc atgggatgat aggttaagct
     2821 gtcttacggt cttttgcccc tgatttttct atggttagaa ttaacgaaaa ttacctcaaa
     2881 ctcgcacctc gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag
     2941 aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag
     3001 cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt
     3061 gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga
     3121 atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc
     3181 aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcccc
     3241 tacgtgctgc tgaagttgcc cgcaacagag agtggaacca accggtgata ccacgatact
     3301 atgactgaga gtcaacgcca tgagcggcct catttcttat tctgagttac aacagtccgc
     3361 accgctgccg gtagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     3421 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgcccaa cagtcccccg
     3481 gccacggggc ctgccaccat acccacgccg aaacaagcgc cctgcaccat tatgttccgg
     3541 atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag
     3601 cgctaaccgt ttttatcagg ctctgggagg cagaataaat gatcatatcg tcaattatta
     3661 cctccacggg gagagcctga gcaaactggc ctcaggcatt tgagaagcac acggtcacac
     3721 tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac
     3781 cctgccctga accgacgacc gggtcgaatt tgctttcgaa tttctgccat tcatccgctt
     3841 attatcactt attcaggcgt agcaaccagg cgtttaaggg
//
