LOCUS       sequence-449896-        5419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn libraries Himar variant GEN backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5419)
  TITLE     Cultivarium plasmids
REFERENCE   2  (bases 1 to 5419)
  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_8
FEATURES             Location/Qualifiers
     source          1..5419
                     /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..3150
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     complement(3490..3506)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3490..3506)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3503..3525)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      3696..4084
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(4217..4241)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(4341..5132,5133..5201))
                     /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     4964..4983
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5202..5306)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(5405..5419,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5419,1..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 tctcaaatgt tacgcagcag caacgatgtt
     2641 acgcagcagg gcagtcgccc taaaacaaag ttaggtggct caagtatggg catcattcgc
     2701 acatgtaggc tcggccctga ccaagtcaaa tccatgcggg ctgctcttga tcttttcggt
     2761 cgtgagttcg gagacgtagc cacctactcc caacatcagc cggactccga ttacctcggg
     2821 aacttgctcc gtagtaagac attcatcgcg cttgctgcct tcgaccaaga agcggttgtt
     2881 ggcgctctcg cggcttacgt tctgcccaag tttgagcagc cgcgtagtga gatctatatc
     2941 tatgatctcg cagtctccgg agagcaccgg aggcagggca ttgccaccgc gctcatcaat
     3001 ctcctcaagc atgaggccaa cgcgcttggt gcttatgtga tctacgtgca agcagattac
     3061 ggtgacgatc ccgcagtggc tctctataca aagttgggca tacgggaaga agtgatgcac
     3121 tttgatatcg acccaagtac cgccacctaa gcttgagcgc atgctcgagt acttcgcctg
     3181 gaccatgctc gccgtcgtct tcggcttcct cctcttcgtc aacctcgcct acgtccccct
     3241 cggccactgg gccgaggcct tcgccggctt cttcaaattt tcagggctgc cgcaccccat
     3301 cgactggggg cttagaccgg ggacttatca tccaacctgt tacgctagca tgaagagcac
     3361 cccaggatac atagattacc acaactccga gcccttccac ctagcgtgct aaccacgcac
     3421 gctattgttg tataggctag gtggaggctc agtgatgata agtctgcgat ggtggatgca
     3481 tgtgtcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcagag ggcacaatcc
     3541 tattccgcgc tatccgacaa tctccaagac attaggtgga gttcagttcg gcgtatggca
     3601 tatgtcgctg gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3661 gccgcgttgc tggcgttaca gctgggcgcg ccccatgtca gccgttaagt gttcctgtgt
     3721 cactcaaaat tgctttgaga ggctctaagg gcttctcagt gcgttacatc cctggcttgt
     3781 tgtccacaac cgttaaacct taaaggcttt aaaagcctta tatattcttt tttttcttat
     3841 aaaacttaaa accttagagg ctatttaagt tgctgattta tattaatttt attgttcaaa
     3901 catgagagct tagtacgtga aacatgagag cttagtacgt tagccatgag agcttagtac
     3961 gttagccatg agggtttagt tcgttaaaca tgagagctta gtacgttaaa catgagagct
     4021 tagtacgtga aacatgagag cttagtacgt actatcaaca ggttgaactg ctgatcttca
     4081 gatcggcgcg ccggccggcc gaagatcctt tgatcttttc tacggggtct gacgctctat
     4141 tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc aaatcgtcct
     4201 cgtgatacca attcggagcc tgcttttttg tacaaacttg ttgataatgg caattcaagg
     4261 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4321 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4381 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4441 gagggcttac catctggccc cagtgctgca atgataccgc gagagccacg ctcaccggct
     4501 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4561 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4621 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4681 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4741 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4801 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4861 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4921 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4981 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5041 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5101 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5161 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5221 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5281 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccaga tacctgaaac
     5341 aaaacccatc gtacggccaa ggaagtctcc aataactgtg atccaccaca agcgccaggg
     5401 ttttcccagt cacgacgtt
//
