LOCUS       sequence-455441-        5356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrates KanR into NS2, 1250 bp homology, two homologous HIP1 
            sites in each of the right and left homology arms.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5356)
  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 5356)
  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: pCMK195
FEATURES             Location/Qualifiers
     source          1..5356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1085..1673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1574..1593
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3169..3963
                     /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(3223..3242)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3833..3852
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      3994..4088
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 gcacctcgct aacggattca ccactccaag aattggagcc aatcaattct tgcggagaac
       61 tgtgaatgcg caaaccaacc cttggcagaa catatccatc gcgtccgcca tctccagcag
      121 ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca cgggtgcgca tgatcgtgct
      181 cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc
      241 accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac
      301 aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcccctac
      361 gtgctgctga agttgcccgc aacagagagt ggaaccaacc ggtgatacca cgatactatg
      421 actgagagtc aacgccatga gcggcctcat ttcttattct gagttacaac agtccgcacc
      481 gctgccggta gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
      541 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgcccaacag tcccccggcc
      601 acggggcctg ccaccatacc cacgccgaaa caagcgccct gcaccattat gttccggatc
      661 tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc
      721 taaccgtttt tatcaggctc tgggaggcag aataaatgat catatcgtca attattacct
      781 ccacggggag agcctgagca aactggcctc aggcatttga gaagcacacg gtcacactgc
      841 ttccggtagt caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct
      901 gccctgaacc gacgaccggg tcgaatttgc tttcgaattt ctgccattca tccgcttatt
      961 atcacttatt caggcgtagc aaccaggcgt ttaagggcac caataactgc cttaaaaaaa
     1021 ttacgccccg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1081 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1141 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1201 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     1261 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1321 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1381 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1441 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1501 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1561 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1621 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1681 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     1741 ctgcgttatc ccctgattct gtggataacc gtattacaat cctgtttgag caactcatag
     1801 gtgacgttgt tgtcggggga ttcgccgctg tcttcgagta agtccattaa ctcggtatct
     1861 tgattatcgc caacccggac atcgagggag attgggtgcc gggaaacacg gaaatattca
     1921 cggatttggg ccggctctaa ttcgagggct tcggcgattt cagcgatggt ggccgcccgg
     1981 cctaggtctt gggccagttg ccgctgggtt ttcttaattt tgttgagctt ttcggtaatg
     2041 tggatcggga gtcgaatcgt gcgaccctgt tgggcgatcg cccgggtgat cgcctggcga
     2101 atccaccagt aggcataggt ggagaactta tacccttttg tggggtcaaa cttctcgaca
     2161 ccccgttcta aaccaaggct gccttcttgg atcaaatcta agaattctag gttgcgcttt
     2221 tgatatttct tggcgatcgc caccacaagc cggagatttg cttcaatcat cttgcgcttt
     2281 gcccgttccc cgatttggag atcccgttgg agttgcttcg gtgtcagttg agcggcctcg
     2341 gcccattctt cacgggttgc ttcgcgatca agggattctt ctagggtttg gcgtttttcc
     2401 tggagggcca tcaggcgctg aacctgtttc ccatagacaa tttcttcttc gtgggttaac
     2461 aagggcaccc gaccgatttc ccggagatag gtacgcacca tatccgcaga acttttgccg
     2521 ttattggggg cttgttgaag atttacttta ggcatttacg ctttaactcc ttaaatctaa
     2581 agaacgaaga actatatggt gcggcccagg ggggcagata ctttacaaaa caatcatcat
     2641 gtaggtcaca atctagaaca caaagaattt ttctaaaaaa taaacgacat accctttaac
     2701 tttcttacgc tttataactt ttaattgagc gctcactact ttagacgggc aacaatcgaa
     2761 aaaggttcaa cgattcatta aaatctcagg gatggggaat tcatcggcgt tctatactta
     2821 attctccttg attgaccaag agcttgaaaa ggggaaaaga tgcgggatta ccgaacttgg
     2881 aatatcgaca gactcaatta gaaatatagt gtcgttgcaa tatgttattc aatgcgataa
     2941 gggtggccct tgcggggaag acatctctca aaaatgagac ctctattgca gtagaggtac
     3001 agggagatgc ccataaatta agacttgacg tcggaattgc cagctggggc gccctctggt
     3061 aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg
     3121 cgcaggggat catgatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3181 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3241 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3301 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3361 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3421 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3481 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3541 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3601 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3661 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3721 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3781 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3841 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3901 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3961 tgagcgggac tctggggttc gagagctcgc ttggactcct gttgatagat ccagtaatga
     4021 cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg
     4081 gtgagaatcc aagcactagt cccttgtgat gatcactacc gtttaaaaga ttgaaaaaga
     4141 atattttttt gaccaataag tccagtttat tcaaaaaaag gcgctttgta tacctttagg
     4201 aaaaagataa tctcagcaaa aataagggca tttaggccag aaatgaagaa cgtagaagta
     4261 ttttggcaag ccagggattt cacctaaaaa ttaacggttg ataatccttc tcaaaaataa
     4321 aagaatgtcc ttgttagtcc aggaaaccga ggtggcgatc gcctttggat caagatctat
     4381 tcccagagtt gcagaatttg tgccgtgatg ggatggcgtt tcgatttttg taacccggtg
     4441 cgatggattg gtcaggtgag tagcgatgga taatttttcc ggcctcttca ccccaaaaag
     4501 atctagcggc ttccctgggg catgggatga taggttaagc tgtcttacgg tcttttgccc
     4561 ctgatttttc tatggttaga attaacgaaa attacctcaa acttaaagca ggttatctct
     4621 ttcccgaaat tgcccggcga gtgaatgggt ttctagcaga aaaccccaat gcgcccatta
     4681 tcaaattagg gattggtgat gtcaccgaac cattacccgc tgcctgtcga gaagcgatgg
     4741 caaaggccat tgacgacatg ggcgatcgcg ccaactttaa agggtatggc cctgaacaag
     4801 gctatgcttg gctacgggaa aaaattgccg cccacgactt ccaggcccgc ggttgcgaca
     4861 ttgacgcctc tgaaattttt gtgtctgatg gtgccaaatg cgataccggg aacatccttg
     4921 atatctttgg caaagacaat accatcgccg tcactgaccc tgtttacccc gtttacgttg
     4981 ataccaacgt gatggcagga cataccggag aagccgacga gtctggtaaa tatggtggcc
     5041 tgacttacat ccccatcacc gctgacaatg actttgtcgc ccaaatcccc acagaaaaag
     5101 tcgatctgat ctatctctgt tttcccaata atcccaccgg ggcaacggcg acgaaagaac
     5161 agctccaagc ttgggtggat tatgccaaga ccaatggttc aattattttc tttgacgctg
     5221 cctatgaagc gttcattacc gacgaaagcc taccccactc gatctacgaa ctagaaggag
     5281 cgaaggattg cgccatcgaa ttccgttctt tctctaaaaa tgcaggcttt acggggactc
     5341 ggtgtgcctt tacctt
//
