LOCUS       sequence-449881-        5590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn libraries Himar variant SPEC backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5590)
  TITLE     Cultivarium plasmids
REFERENCE   2  (bases 1 to 5590)
  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_12
FEATURES             Location/Qualifiers
     source          1..5590
                     /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)"
     primer_bind     complement(3661..3677)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3661..3677)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3674..3696)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      3867..4255
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(4388..4412)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(4512..5303,5304..5372))
                     /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     5135..5154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5373..5477)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(5576..5590,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5590,1..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 tgaatacata cgaacaaatt
     2641 aataaagtga aaaaaatact tcggaaacat ttaaaaaata accttattgg tacttacatg
     2701 tttggatcag gagttgagag tggactaaaa ccaaatagtg atcttgactt tttagtcgtc
     2761 gtatctgaac cattgacaga tcaaagtaaa gaaatactta tacaaaaaat tagacctatt
     2821 tcaaagaaaa taggagataa aagcaactta cgatatattg aattaacaat tattattcag
     2881 caagaaatgg taccgtggaa tcatcctccc aaacaagaat ttatttatgg agaatggtta
     2941 caagagcttt atgaacaagg atacattcct cagaaggaat taaattcaga tttaaccata
     3001 atgctttacc aagcaaaacg aaaaaataaa agaatatacg gaaattatga cttagaggaa
     3061 ttactacctg atattccatt ttctgatgtg agacgcgcca ttatggattc gtcagaggaa
     3121 ttaatagata attatcagga tgatgaaacc aactctatat taactttatg ccgtatgatt
     3181 ttaactatgg acacgggtaa aatcatacca aaagatattg cgggaaatgc agtggctgaa
     3241 tcttctccat tagaacatag ggagagaatt ttgttagcag ttcgtagtta tcttggagag
     3301 aatattgaat ggacaaacga gaacgtcaac cttactataa attacttaaa taacagatta
     3361 aaaaaattat aagcttaaaa attgaaaaaa tggtggaaac acttttttca atttttttgt
     3421 tttattattt aatatttggg aaatattcat tctaattggt aatcagattt tagaagttta
     3481 aactagaccg gggacttatc atccaacctg ttacgctagc atgaagagca ccccaggata
     3541 catagattac cacaactccg agcccttcca cctagcgtgc taaccacgca cgctattgtt
     3601 gtataggcta ggtggaggct cagtgatgat aagtctgcga tggtggatgc atgtgtcatg
     3661 gtcatagctg tttcctgtgt gaaattgtta tccgctcaga gggcacaatc ctattccgcg
     3721 ctatccgaca atctccaaga cattaggtgg agttcagttc ggcgtatggc atatgtcgct
     3781 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3841 ctggcgttac agctgggcgc gccccatgtc agccgttaag tgttcctgtg tcactcaaaa
     3901 ttgctttgag aggctctaag ggcttctcag tgcgttacat ccctggcttg ttgtccacaa
     3961 ccgttaaacc ttaaaggctt taaaagcctt atatattctt ttttttctta taaaacttaa
     4021 aaccttagag gctatttaag ttgctgattt atattaattt tattgttcaa acatgagagc
     4081 ttagtacgtg aaacatgaga gcttagtacg ttagccatga gagcttagta cgttagccat
     4141 gagggtttag ttcgttaaac atgagagctt agtacgttaa acatgagagc ttagtacgtg
     4201 aaacatgaga gcttagtacg tactatcaac aggttgaact gctgatcttc agatcggcgc
     4261 gccggccggc cgaagatcct ttgatctttt ctacggggtc tgacgctcta ttcaacaaag
     4321 ccgccgtccc gtcaagtcag cgtaaatggg tagggggctt caaatcgtcc tcgtgatacc
     4381 aattcggagc ctgctttttt gtacaaactt gttgataatg gcaattcaag gatcttcacc
     4441 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4501 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4561 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4621 ccatctggcc ccagtgctgc aatgataccg cgagagccac gctcaccggc tccagattta
     4681 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4741 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4801 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4861 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4921 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4981 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5041 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5101 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5161 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5221 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5281 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5341 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5401 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5461 caaatagggg ttccgcgcac atttccccga aaagtgccag atacctgaaa caaaacccat
     5521 cgtacggcca aggaagtctc caataactgt gatccaccac aagcgccagg gttttcccag
     5581 tcacgacgtt
//
