LOCUS       sequence-449895-        5701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn libraries Himar variant KAN backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5701)
  TITLE     Cultivarium plasmids
REFERENCE   2  (bases 1 to 5701)
  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"
COMMENT     Sequence Label: pGLU_7
FEATURES             Location/Qualifiers
     source          1..5701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            161..269
                     /note="incP origin of transfer"
     RBS             1797..1808
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2617..3432
                     /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             2617..3432
                     /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     complement(2690..2709)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(3772..3788)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3772..3788)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3785..3807)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      3978..4366
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(4499..4523)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(4623..5414,5415..5483))
                     /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     5246..5265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5484..5588)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(5687..5701,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5701,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtcat gcataatccg cacgcatctg gaataaggaa gtgccattcc
       61 gcctgacctt agcgtgcgaa cagcacgcta ttgttgtatc attactcgca tccattctca
      121 ggctgtctcg tctcgtctcc gagctcttca atgaggagta cggccagcct cgcagagcag
      181 gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg
      241 cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt
      301 ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa
      361 aaatcgctat aatgaccccg aagcagggtt atgcagcgga aaagtcgttg ggattgtcct
      421 gaccgaagcg ggacaggtcc acgagcaggc aatgcttgtt gggcatggtg aggtggatgt
      481 ctttcacttc cggcacgcag ccgaggactt tttgcgccat atcgtaaagt gtgtgctgaa
      541 cggattcgct ttcgtgttca gcgaaagagg cgagaaggag attccaagct tattcaacat
      601 agttcccttc aagagcgata caacgattat aacgaccttc caattttttg ataccatttt
      661 ggtagtactc cttcggtttt gcctcaaaat aggcctcagt ttcggcgatc acctcttcat
      721 tgcagccaaa ttttttccct gcgagcatcc ttttgaggtc tgagaacaag aaaaagtcgc
      781 tgggggccag atctggagaa tacggcgggt ggggaagcaa ttcgaagccc aattcatgaa
      841 tttttgccat cgttctcaat gacttgtggc acggtgcgtt gtcttggtgg aacaacactt
      901 ttttcttctt catgtggggc cgttttgccg cgatttcgac cttcaaacgc tccaataacg
      961 ccatataata gtcactgttg atggtttttc ccttctcaag ataatcgata aaaattattc
     1021 catgcgcatc ccaaaaaaca gaggccatta ctttgccagc ggacttttga gtctttccac
     1081 gcttcggaga cggttcaccg gtcgctgtcc actcagccga ctgtcgattg gactcaggag
     1141 tgtagtgatg gagccatgtt tcatccattg tcacatatcg acggaaaaac tcgggtgtat
     1201 tacgagttaa cagctgcaaa caccgcttag aatcatcaac acgtcgttgt ttttggtcaa
     1261 atgtgagctc gcgcggcacc catttcgcac agagcttccg catatccaaa tattgatgaa
     1321 tgatatgacc aacacgttcc tttgatatct ttaaggcctc tgctatctcg atcaacttca
     1381 ttttacggtc attcaaaatc attttgtgga tttttttgat gttttcgtcg gtaaccacct
     1441 ctttcgggcg tccactgcgt tcaccgtcct ccgtgctcat ttcaccacgc ttgaattttg
     1501 cataccaatc aattattgtt gatttccctg gggcagagtc cggaaactca ttatcaagcc
     1561 aagtttttgc ttccactgta ttttttccct tcagaaaaca gtattttatc aaaacacgaa
     1621 attccttttt ttccattgag acccagtcga aagtgaaacg tgatttcatg cgtcattttg
     1681 aacattttgt aaatcttatt taataatgtg tgcggcaatt cacatttaat ttatgaatgt
     1741 tttcttaaca tcgcggcaac tcaagaaacg gcaggttcgg atcttagcta ctagagaaag
     1801 aggagaaata ctagatggtg agtaagggtg aagccgtgat taaagagttc atgcgcttca
     1861 aagtgcatat ggagggctca atgaacggcc atgaatttga aattgaaggt gaaggagaag
     1921 gccgtccgta tgagggcacc cagaccgcca aactgaaagt cacaaagggt ggcccgctgc
     1981 cgttttcgtg ggatatttta tcccctcaat tcatgtacgg cagccgggcg tttattaaac
     2041 acccggcgga tatcccggat tactataagc aatcttttcc agaaggcttt aaatgggagc
     2101 gcgtgatgaa ttttgaagac ggaggtgctg ttaccgtgac ccaggacacg agcttagaag
     2161 acggtacctt gatctataag gtaaagctgc gtggtaccaa tttcccgccg gatgggccag
     2221 ttatgcagaa aaagactatg ggctgggaag catcgacgga gcgtctttac cccgaagacg
     2281 gcgtcctgaa aggcgatatt aaaatggccc tgcgactgaa agacggcggt cgctaccttg
     2341 cagatttcaa aacaacgtat aaagcgaaaa aacctgtgca gatgcccggg gcgtacaacg
     2401 ttgatcgcaa actagatatc acttcacaca acgaagatta cacggtagtc gaacagtatg
     2461 aacgtagcga ggggcgccat agcaccggag gcatggatga actctataaa taattcagcc
     2521 aaaaaactta agaccgccgg tcttgtccac taccttgcag taatgcggtg gacaggatcg
     2581 gcggttttct tttctcttct caagactggg tctcaaatga gccatattca acgggaaacg
     2641 tcttgctcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     2701 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat
     2761 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     2821 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     2881 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     2941 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     3001 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     3061 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     3121 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     3181 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     3241 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     3301 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     3361 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     3421 gagtttttct aagcttgagc gcatgctcga gtacttcgcc tggaccatgc tcgccgtcgt
     3481 cttcggcttc ctcctcttcg tcaacctcgc ctacgtcccc ctcggccact gggccgaggc
     3541 cttcgccggc ttcttcaaat tttcagggct gccgcacccc atcgactggg ggcttagacc
     3601 ggggacttat catccaacct gttacgctag catgaagagc accccaggat acatagatta
     3661 ccacaactcc gagcccttcc acctagcgtg ctaaccacgc acgctattgt tgtataggct
     3721 aggtggaggc tcagtgatga taagtctgcg atggtggatg catgtgtcat ggtcatagct
     3781 gtttcctgtg tgaaattgtt atccgctcag agggcacaat cctattccgc gctatccgac
     3841 aatctccaag acattaggtg gagttcagtt cggcgtatgg catatgtcgc tggaaagaac
     3901 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgtta
     3961 cagctgggcg cgccccatgt cagccgttaa gtgttcctgt gtcactcaaa attgctttga
     4021 gaggctctaa gggcttctca gtgcgttaca tccctggctt gttgtccaca accgttaaac
     4081 cttaaaggct ttaaaagcct tatatattct tttttttctt ataaaactta aaaccttaga
     4141 ggctatttaa gttgctgatt tatattaatt ttattgttca aacatgagag cttagtacgt
     4201 gaaacatgag agcttagtac gttagccatg agagcttagt acgttagcca tgagggttta
     4261 gttcgttaaa catgagagct tagtacgtta aacatgagag cttagtacgt gaaacatgag
     4321 agcttagtac gtactatcaa caggttgaac tgctgatctt cagatcggcg cgccggccgg
     4381 ccgaagatcc tttgatcttt tctacggggt ctgacgctct attcaacaaa gccgccgtcc
     4441 cgtcaagtca gcgtaaatgg gtagggggct tcaaatcgtc ctcgtgatac caattcggag
     4501 cctgcttttt tgtacaaact tgttgataat ggcaattcaa ggatcttcac ctagatcctt
     4561 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4621 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4681 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4741 cccagtgctg caatgatacc gcgagagcca cgctcaccgg ctccagattt atcagcaata
     4801 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4861 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4921 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4981 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5041 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5101 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5161 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5221 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5281 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5341 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5401 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5461 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5521 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5581 gttccgcgca catttccccg aaaagtgcca gatacctgaa acaaaaccca tcgtacggcc
     5641 aaggaagtct ccaataactg tgatccacca caagcgccag ggttttccca gtcacgacgt
     5701 t
//
