LOCUS       sequence-449888-        6073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn libraries Tn5 variant KAN backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6073)
  TITLE     Cultivarium plasmids
REFERENCE   2  (bases 1 to 6073)
  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_128
FEATURES             Location/Qualifiers
     source          1..6073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(144..160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(157..179)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      350..738
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(871..895)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(995..1786,1787..1855))
                     /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     1618..1637
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1856..1960)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2059..2081
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2073..2090
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2074..2090
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            2234..2342
                     /note="incP origin of transfer"
     CDS             complement(2666..4093)
                     /codon_start=1
                     /product="transposase from the bacterial Tn5 transposon 
                     (Reznikoff, 1993)"
                     /label=Tn5 transposase
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     RBS             4254..4265
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5073..5888
                     /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             5073..5888
                     /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(5146..5165)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    6051..6069
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
ORIGIN
        1 agcatgaaga gcaccccagg atacatagat taccacaact ccgagccctt ccacctagcg
       61 tgctaaccac gcacgctatt gttgtatagg ctaggtggag gctcagtgat gataagtctg
      121 cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
      181 agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg tggagttcag
      241 ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc aaaaggccag caaaaggcca
      301 ggaaccgtaa aaaggccgcg ttgctggcgt tacagctggg cgcgccccat gtcagccgtt
      361 aagtgttcct gtgtcactca aaattgcttt gagaggctct aagggcttct cagtgcgtta
      421 catccctggc ttgttgtcca caaccgttaa accttaaagg ctttaaaagc cttatatatt
      481 cttttttttc ttataaaact taaaacctta gaggctattt aagttgctga tttatattaa
      541 ttttattgtt caaacatgag agcttagtac gtgaaacatg agagcttagt acgttagcca
      601 tgagagctta gtacgttagc catgagggtt tagttcgtta aacatgagag cttagtacgt
      661 taaacatgag agcttagtac gtgaaacatg agagcttagt acgtactatc aacaggttga
      721 actgctgatc ttcagatcgg cgcgccggcc ggccgaagat cctttgatct tttctacggg
      781 gtctgacgct ctattcaaca aagccgccgt cccgtcaagt cagcgtaaat gggtaggggg
      841 cttcaaatcg tcctcgtgat accaattcgg agcctgcttt tttgtacaaa cttgttgata
      901 atggcaattc aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      961 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1021 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1081 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagagc
     1141 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     1201 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     1261 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     1321 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     1381 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     1441 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1501 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1561 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1621 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1681 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1741 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1801 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1861 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1921 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1981 cagatacctg aaacaaaacc catcgtacgg ccaaggaagt ctccaataac tgtgatccac
     2041 cacaagcgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt catgcataat
     2101 ccgcacgcat ctggaataag gaagtgccat tccgcctgac cttagcgtgc gaacagcacg
     2161 ctattgttgt atcattactc gcatccattc tcaggctgtc tcgtctcgtc tccgagctct
     2221 tcaatgagga gtacggccag cctcgcagag caggattccc gttgagcacc gccaggtgcg
     2281 aataagggac agtgaagaag gaacacccgc tcgcgggtgg gcctacttca cctatcctgc
     2341 ccggctgacg ccgttggata caccaaggaa agtctacacg aaccctttgg caaaatcctg
     2401 tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc tataatgacc ccgaagcagg
     2461 gttatgcagc ggaaaagtcg ttgggattgt cctgaccgaa gcgggacagg tccacgagca
     2521 ggcaatgctt gttgggcatg gtgaggtgga tgtctttcac ttccggcacg cagccgagga
     2581 ctttttgcgc catatcgtaa agtgtgtgct gaacggattc gctttcgtgt tcagcgaaag
     2641 aggcgagaag gagattccaa gcttagattt taatgccctg cgccatcagg tctttcgcgg
     2701 ccagaaagcc atccagtttg ctttgcagcg cttcccaacc ttcccacagc gcaccccagc
     2761 tcgcaatgcc ggtacgtttg ctatccataa agccgcccag acgcgcaatc gccatatacg
     2821 cccattgcag gctgcccgct ttttctttgc gtttgcgttt gcctttatcc agatagccca
     2881 gcagttggca ttcatccggg gtcagcacgg tttccgcgct ctggctttca acgtgttccg
     2941 cttctttcag caggccctgc gcacgcagtg cttgcggcgg agtaaaagat tcacgcagtt
     3001 gcagcagacg caccgccaca aagctcagaa tgctcaccat acgttccagg ttatccggtt
     3061 cttccatacg ctgacgttcc gcacccgcac ccgttttcca cgctttgtga aattcttcaa
     3121 tgcgccaacg atgggtataa atatcaatca cacgcagcgc ttgggccaga ctttccaccg
     3181 gctcgctggt cagcagcagc catttcagcg gggtttcgcc tttcggcgga ttaatttctt
     3241 cggccagcac cgcgttcagg gtaatgttgc cctgtttcag ggtaatacgg ccgctacgca
     3301 ggctcaggct cgctttacgc gccggacggt ttttacgttt gccacgttta tccaccacgc
     3361 ctttctgcgg aatgctaatc tgatagccgc ccagttccgg ctggtttttc aggtgatcat
     3421 acagatacag gccgctttcc acatctttac gcggatgttt gctacgcacc acaaaacgtt
     3481 cgttatgggc cagtttatct tgcagatacg catgaatatc cgcttcacga tcgcacaccg
     3541 caatcacgtt gctcatcatg ctgcccattc tcagacgcga agttgcagca gcggccagcc
     3601 atttgccgct ttctttttca tccgcatccg ccggatcatc cggacgcatc caccattctt
     3661 gatgcagcag gcccacggta cgaaaggtgg tcgcttccag cagcagcacg ctatgcaccc
     3721 accaaccacg gcttttatcc tgaatgctac ccagtttgcc cagttcttcc gccacctgat
     3781 gacgatagct cagagaggtg gtatcttcaa ttgccagcag ttccggaaat tcctgggcca
     3841 gtttcacggt ctgcatggca cccgctttac gaatcgcttc cgcgctcacg ttcggattac
     3901 gaataaaacg atacgcgcct tcctgcatgg ctttgctgcc ttcgctgcta atggtaatgc
     3961 ttttgccgct atatttggcc agttgcgccg caacattcac cagacgcgcg gtacgacgcg
     4021 gatcacccag cgcagcacta gaaaacacgc ttttcgccca atccgccgca cgatgcagtg
     4081 cactggtaat cattgagacc cagtcgaaag tgaaacgtga tttcatgcgt cattttgaac
     4141 attttgtaaa tcttatttaa taatgtgtgc ggcaattcac atttaattta tgaatgtttt
     4201 cttaacatcg cggcaactca agaaacggca ggttcggatc ttagctacta gagaaagagg
     4261 agaaatacta gatggtgagt aagggtgaag ccgtgattaa agagttcatg cgcttcaaag
     4321 tgcatatgga gggctcaatg aacggccatg aatttgaaat tgaaggtgaa ggagaaggcc
     4381 gtccgtatga gggcacccag accgccaaac tgaaagtcac aaagggtggc ccgctgccgt
     4441 tttcgtggga tattttatcc cctcaattca tgtacggcag ccgggcgttt attaaacacc
     4501 cggcggatat cccggattac tataagcaat cttttccaga aggctttaaa tgggagcgcg
     4561 tgatgaattt tgaagacgga ggtgctgtta ccgtgaccca ggacacgagc ttagaagacg
     4621 gtaccttgat ctataaggta aagctgcgtg gtaccaattt cccgccggat gggccagtta
     4681 tgcagaaaaa gactatgggc tgggaagcat cgacggagcg tctttacccc gaagacggcg
     4741 tcctgaaagg cgatattaaa atggccctgc gactgaaaga cggcggtcgc taccttgcag
     4801 atttcaaaac aacgtataaa gcgaaaaaac ctgtgcagat gcccggggcg tacaacgttg
     4861 atcgcaaact agatatcact tcacacaacg aagattacac ggtagtcgaa cagtatgaac
     4921 gtagcgaggg gcgccatagc accggaggca tggatgaact ctataaataa ttcagcaaaa
     4981 aacttaagac cgccggtctt gtccactacc ttgcagtaat gcggtggaca ggatcggcgg
     5041 ttttcttttc tcttctcaag actgggtctc aaatgagcca tattcaacgg gaaacgtctt
     5101 gctcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     5161 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     5221 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     5281 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     5341 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
     5401 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     5461 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     5521 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     5581 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     5641 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     5701 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     5761 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     5821 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     5881 ttttctaagc ttgagcgcat gctcgagtac ttcgcctgga ccatgctcgc cgtcgtcttc
     5941 ggcttcctcc tcttcgtcaa cctcgcctac gtccccctcg gccactgggc cgaggccttc
     6001 gccggcttct tcaaattttc agggctgccg caccccatcg actgggggct agatgtgtat
     6061 aagagacagc gct
//
