LOCUS       sequence-461200-        6155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tn libraries Himar variant TET backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6155)
  TITLE     Cultivarium plasmids
REFERENCE   2  (bases 1 to 6155)
  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_10
FEATURES             Location/Qualifiers
     source          1..6155
                     /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..3993
                     /codon_start=1
                     /gene="Bacillus subtilis tetL"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
                     KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
                     LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
                     HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
                     ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
                     MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
                     SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
                     GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
                     "
     primer_bind     complement(4226..4242)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4226..4242)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4239..4261)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      4432..4820
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(4953..4977)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(5077..5868,5869..5937))
                     /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     5700..5719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5938..6042)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(6141..6155,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6155,1..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 atacatccta ttcacaatcg
     2641 aatttacgac acaaccaaat tttaatttgg ctttgcattt tatctttttt tagcgtatta
     2701 aatgaaatgg ttttgaacgt cagcttacct gatattgcaa atgattttaa taaacctcct
     2761 gcgagtacaa actgggtgaa cacagccttt atgttaacct tttccattgg aacagctgta
     2821 tatggaaagc tatctgatca attaggcatc aaaaggttac tcctatttgg aattataata
     2881 aattgtttcg ggtcggtaat tgggtttgtt ggccattctt tcttttcctt acttattatg
     2941 gctcgtttta ttcaaggggc tggtgcagct gcatttccag cactcgtaat ggttgtagtt
     3001 gcgcgctata ttccaaagga aaataggggt aaagcatttg gtcttattgg atcgatagta
     3061 gccatgggag aaggagtcgg tccagcgatt ggtggaatga tagcccatta tattcattgg
     3121 tcctatcttc tactcattcc tatgataaca attatcactg ttccgtttct tatgaaatta
     3181 ttaaagaaag aagtaaggat aaaaggtcat tttgatatca aaggaattat actaatgtct
     3241 gtaggcattg tattttttat gttgtttaca acatcatata gcatttcttt tcttatcgtt
     3301 agcgtgctgt cattcctgat atttgtaaaa catatcagga aagtaacaga tccttttgtt
     3361 gatcccggat tagggaaaaa tatacctttt atgattggag ttctttgtgg gggaattata
     3421 tttggaacag tagcagggtt tgtctctatg gttccttata tgatgaaaga tgttcaccag
     3481 ctaagtactg ccgaaatcgg aagtgtaatt attttccctg gaacaatgag tgtcattatt
     3541 ttcggctaca ttggtgggat acttgttgat agaagaggtc ctttatacgt gttaaacatc
     3601 ggagttacat ttctttctgt tagcttttta actgcttcct ttcttttaga aacaacatca
     3661 tggttcatga caattataat cgtatttgtt ttaggtgggc tttcgttcac caaaacagtt
     3721 atatcaacaa ttgtttcaag tagcttgaaa cagcaggaag ctggtgctgg aatgagtttg
     3781 cttaacttta ccagcttttt atcagaggga acaggtattg caattgtagg tggtttatta
     3841 tccataccct tacttgatca aaggttgtta cctatggaag ttgatcagtc aacttatctg
     3901 tatagtaatt tgttattact tttttcagga atcattgtca ttagttggct ggttaccttg
     3961 aatgtatata aacattctca aagggatttc taagcttatc gttaagggat caactttggg
     4021 agagagttca aaattgatcc tttttttata gaccggggac ttatcatcca acctgttacg
     4081 ctagcatgaa gagcacccca ggatacatag attaccacaa ctccgagccc ttccacctag
     4141 cgtgctaacc acgcacgcta ttgttgtata ggctaggtgg aggctcagtg atgataagtc
     4201 tgcgatggtg gatgcatgtg tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4261 tcagagggca caatcctatt ccgcgctatc cgacaatctc caagacatta ggtggagttc
     4321 agttcggcgt atggcatatg tcgctggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4381 caggaaccgt aaaaaggccg cgttgctggc gttacagctg ggcgcgcccc atgtcagccg
     4441 ttaagtgttc ctgtgtcact caaaattgct ttgagaggct ctaagggctt ctcagtgcgt
     4501 tacatccctg gcttgttgtc cacaaccgtt aaaccttaaa ggctttaaaa gccttatata
     4561 ttcttttttt tcttataaaa cttaaaacct tagaggctat ttaagttgct gatttatatt
     4621 aattttattg ttcaaacatg agagcttagt acgtgaaaca tgagagctta gtacgttagc
     4681 catgagagct tagtacgtta gccatgaggg tttagttcgt taaacatgag agcttagtac
     4741 gttaaacatg agagcttagt acgtgaaaca tgagagctta gtacgtacta tcaacaggtt
     4801 gaactgctga tcttcagatc ggcgcgccgg ccggccgaag atcctttgat cttttctacg
     4861 gggtctgacg ctctattcaa caaagccgcc gtcccgtcaa gtcagcgtaa atgggtaggg
     4921 ggcttcaaat cgtcctcgtg ataccaattc ggagcctgct tttttgtaca aacttgttga
     4981 taatggcaat tcaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5041 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5101 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5161 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5221 gccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5281 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5341 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5401 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5461 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5521 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5581 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5641 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5701 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5761 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5821 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5881 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5941 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6001 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6061 gccagatacc tgaaacaaaa cccatcgtac ggccaaggaa gtctccaata actgtgatcc
     6121 accacaagcg ccagggtttt cccagtcacg acgtt
//
