LOCUS       sequence-455428-        4685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrates slr0214 (HIP1 methyltransferase) into gidB-atpI locus of 
            E. coli DH10B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4685)
  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 4685)
  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: pCMK83
FEATURES             Location/Qualifiers
     source          1..4685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(231..248)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(365..910)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(445..464)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2458..2469
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(join(2984..2989,2990..3006,3007..3012))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        3647..3751
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3752..3820,3821..4612)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3970..3989)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
       61 gagcgtcaga ccccgtagaa aagatcaaag gatcttcgcg ctagcggagt gtatactggc
      121 ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag gagaaaaaag
      181 gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc ctcgctcact
      241 gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga
      301 gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc
      361 cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt
      421 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg
      481 tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt
      541 tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt
      601 atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      661 tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga
      721 ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga
      781 ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga
      841 aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa
      901 acgatctcaa gaagatcatc ttattaatca gataaaatat ttctagattt cagtgcaatt
      961 tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt aaacgccagc
     1021 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     1081 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc ctgaagccca
     1141 tttcactctg ttggatagcc ttggtaaacg cgtgcgtttc cttcgtcagg tgcaacatga
     1201 gcttaaactg gagaatattg aaccagtaca gagcagggta gaagagtttc cttcagagcc
     1261 gccatttgat ggcgtaatta gccgcgcttt tgcctctctg aacgatatgg tgagctggtg
     1321 ccaccatctt cctggtgagc aaggccgttt ctacgcgctg aaagggcaaa tgccggaaga
     1381 tgaaatcgct ttgttgcccg aagaatatca ggtcgaatca gtggttaaac ttcaggttcc
     1441 agccctggat ggcgaacgtc atctggtggt gattaaagca aataaaattt aatttttatc
     1501 aaaaaaatca taaaaaattg accggttaga ctgttaacaa caaccaggtt ttctactgat
     1561 ataactggtt acatttaacg ccacgttcac tcttttgcat caacaagata acgtggcttt
     1621 ttttggtaag cagaaaataa gtcattagtg aaaatatggc taactgcttt aggaatggga
     1681 tttggcacat aaatatcgac tagcctgggg caaattgtaa tgtagtaatg ttcggtcttg
     1741 gggctccagt tcataaaccg gcaaatcctc aacagaacaa ttcaatgcgg ccataacctg
     1801 ttctcccaaa accttagcca gcaaaggtac gacggagtta ccaatttccc gaaagccgtg
     1861 ccagatggtg ttgtggaact gaaaccaatc cggataggta tggagtctgg ccgcttctct
     1921 cacagttata catctaggta aggcgtagtg tattggccta ggggcggtat aagcaccctt
     1981 atcactagca gtgccggccc tcagggtatt acataaacca gtttttgata gccgcagaaa
     2041 gcgactggtt ctttccaccg ttccagggac agtatccgca aatcttttca ttgatttttc
     2101 agtatgttga gaaccaatat ggccataaac taaattatta acttggcgct tatggcagag
     2161 ggaaaatgct ccatttgcat ggatggcata actttttctc agggctgaat aatccaataa
     2221 ttctgatgga ataccttggt cttctcctac tgcatagggt gtatggtcaa ctagatcatc
     2281 aatggcttct ccacaggtaa atggcggttt ataagctatg ttagaaactg atttttcaga
     2341 acaattactt tgttttaatt cagcaacgtc aaaatagagt ggaaggggat aggtgacagg
     2401 cttgacatct tttctggagc cgagaataat caaccttttt cgtttttggg gagcaccata
     2461 gagggacgca tctagcacta aaattggttt ttcgatgtgg taaccaatgg tttcaaattc
     2521 cctaactaat tcctctaaaa attttttatg ttgccccgat cgcataccgg gaacattttc
     2581 aaataaaaaa tatttgggtt tgagatcccg aatcattcgc acatattcaa acaccaagga
     2641 gttacgagga tcatccaact gccttttacc catgagggaa aagccttgac agggagggcc
     2701 tccagcaatc aggtctatat cgcttgaata acctttatta tttaactttc ttagaatttc
     2761 gcccgcactt accagagcaa tatcccgaca gatagtaaca ccatagggaa aattatgatg
     2821 atggaccaaa caatgcactg catcaaattc caccgcggcg gcaatatcaa atccagcggc
     2881 ttctaaaccc aaggacatgc caccacagcc cgcaaacaag tcgatggcaa tgggtctggc
     2941 catggttttt cctcctagca atttaactgt gataaactac cgcattaaag cttatcgatg
     3001 ataagctgtc aaacatgaga attacaactt atatcgtatg gggctgacat acttattgtt
     3061 tgaaatcacg ggggcgcacc gtataatttg accgcttttt gatgcttgac tctaagcctt
     3121 aaagaaagtt ttatacgaca cgcggcatac ctcgaaggga gcaggagtga aaaacgtgat
     3181 gtctgtgtcg ctcgtgagtc gaaacgttgc tcggaagctt ctgctcgttc agttactggt
     3241 ggtgatagca agtggattgc tgttcagcct caaagacccc ttctggggcg tctctgcaat
     3301 aagcgggggc ctggcagtct ttctgcctaa cgttttgttt atgatatttg cctggcgtca
     3361 ccaggcgcat acaccagcga aaggccgggt ggcctggaca ttcgcatttg gcgaagcttt
     3421 caaagttctg gcgatgttgg tgttactggt ggtggcgttg gcggttttaa aggcggtatt
     3481 cttgccgctg atcgttacgt gggttttggt gctggtggtt cagatactgg caccggctgt
     3541 aattaacaac aaagggtaac tcgtttcgat acgcctattt ttataggtta atgtcatgat
     3601 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3661 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3721 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     3781 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3841 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3901 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3961 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4021 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4081 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4141 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     4201 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4261 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4321 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4381 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4441 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4501 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4561 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     4621 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4681 ctagg
//
