LOCUS       sequence-92136-e        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MethyloBrick cloning vector, no promoter; TcR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6558)
  AUTHORS   Schada von Borzyskowski L, Remus-Emsermann M, Weishaupt R, Vorholt 
            JA, Erb TJ
  TITLE     A Set of Versatile Brick Vectors and Promoters for the Assembly, 
            Expression, and Integration of Synthetic Operons in Methylobacterium
            extorquens AM1 and Other Alphaproteobacteria.
  JOURNAL   ACS Synth Biol. 2014 Aug 8.
  PUBMED    25105793
REFERENCE   2  (bases 1 to 6558)
  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"; journalName: "ACS Synth 
            Biol. 2014 Aug 8."
COMMENT     Sequence Label: pTE100
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..642
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1594..1613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1845..1862)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1845..1861)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1854..1876)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2070..2089)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2189..2211
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2249..2267)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             3053..4252
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             complement(4483..5631)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(5903..6274)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            6307..6416
                     /note="incP origin of transfer"
ORIGIN
        1 gaccctttcc gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg
       61 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc
      121 gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca
      181 cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc
      241 cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt
      301 tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg
      361 gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca
      421 gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt
      481 gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac
      541 caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc
      601 cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc
      661 ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agccaagctg
      721 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      781 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      841 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      901 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
      961 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1021 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1081 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1141 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1201 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1261 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1321 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1381 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1441 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1501 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1561 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1621 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1681 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1741 ttttgctcac atggaattct gtacatctag aaataagaag gagatataat taatccatgg
     1801 caattgaagc ttagatcttg actagtcctg caggtaccta attcactggc cgtcgtttta
     1861 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     1921 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     1981 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     2041 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2101 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2161 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     2221 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     2281 gtcatgataa taatggtttc ttagcaccct ttctcggtcc ttcaacgttc ctgacaacga
     2341 gcctcctttt cgccaatcca tcgacaatca ccgcgagtcc ctgctcgaac gctgcgtccg
     2401 gaccggcttc gtcgaaggcg tctatcgcgg cccgcaacag cggcgagagc ggagcctgtt
     2461 caacggtgcc gccgcgctcg ccggcatcgc tgtcgccggc ctgctcctca agcacggccc
     2521 caacagtgaa gtagctgatt gtcatcagcg cattgacggc gtccccggcc gaaaaacccg
     2581 cctcgcagag gaagcgaagc tgcgcgtcgg ccgtttccat ctgcggtgcg cccggtcgcg
     2641 tgccggcatg gatgcgcgcg ccatcgcggt aggcgagcag cgcctgcctg aagctgcggg
     2701 cattcccgat cagaaatgag cgccagtcgt cgtcggctct cggcaccgaa tgcgtatgat
     2761 tctccgccag catggcttcg gccagtgcgt cgagcagcgc ccgcttgttc ctgaagtgcc
     2821 agtaaagcgc cggctgctga acccccaacc gttccgccag tttgcgtgtc gtcagaccgt
     2881 ctacgccgac ctcgttcaac aggtccaggg cggcacggat cactgtattc ggctgcaact
     2941 ttgtcatgct tgacacttta tcactgataa acataatatg tccaccaact tatcagtgat
     3001 aaagaatccg cgcgttcaat cggaccagcg gaggctggtc cggaggccag acgtgaaacc
     3061 caacataccc ctgatcgtaa ttctgagcac tgtcgcgctc gacgctgtcg gcatcggcct
     3121 gattatgccg gtgctgccgg gcctcctgcg cgatctggtt cactcgaacg acgtcaccgc
     3181 ccactatggc attctgctgg cgctgtatgc gttggtgcaa tttgcctgcg cacctgtgct
     3241 gggcgcgctg tcggatcgtt tcgggcggcg gccaatcttg ctcgtctcgc tggccggcgc
     3301 cactgtcgac tacgccatca tggcgacagc gcctttcctt tgggttctct atatcgggcg
     3361 gatcgtggcc ggcatcaccg gggcgactgg ggcggtagcc ggcgcttata ttgccgatat
     3421 cactgatggc gatgagcgcg cgcggcactt cggcttcatg agcgcctgtt tcgggttcgg
     3481 gatggtcgcg ggacctgtgc tcggtgggct gatgggcggt ttctcccccc acgctccgtt
     3541 cttcgccgcg gcagccttga acggcctcaa tttcctgacg ggctgtttcc ttttgccgga
     3601 gtcgcacaaa ggcgaacgcc ggccgttacg ccgggaggct ctcaacccgc tcgcttcgtt
     3661 ccggtgggcc cggggcatga ccgtcgtcgc cgccctgatg gcggtcttct tcatcatgca
     3721 acttgtcgga caggtgccgg ccgcgctttg ggtcattttc ggcgaggatc gctttcactg
     3781 ggacgcgacc acgatcggca tttcgcttgc cgcatttggc attctgcatt cactcgccca
     3841 ggcaatgatc accggccctg tagccgcccg gctcggcgaa aggcgggcac tcatgctcgg
     3901 aatgattgcc gacggcacag gctacatcct gcttgccttc gcgacacggg gatggatggc
     3961 gttcccgatc atggtcctgc ttgcttcggg tggcatcgga atgccggcgc tgcaagcaat
     4021 gttgtccagg caggtggatg aggaacgtca ggggcagctg caaggctcac tggcggcgct
     4081 caccagcctg acctcgatcg tcggacccct cctcttcacg gcgatctatg cggcttctat
     4141 aacaacgtgg aacgggtggg catggattgc aggcgctgcc ctctacttgc tctgcctgcc
     4201 ggcgctgcgt cgcgggcttt ggagcggcgc agggcaacga gccgatcgct gatcgtggaa
     4261 acgataggcc tatgccatgc gggtcaaggc gacttccggc aagctatacg cgccctagaa
     4321 ttgtcaattt taatcctctg tttatcggca gttcgtagag cgcgccgtgc gtcccgagcg
     4381 atactgagcg aagcaagtgc gtcgagcagt gcccgcttgt tcctgaaatg ccagtaaagc
     4441 gctggctgct gaacccccag ccggaactga ccccacaagg ccctagcgtt tgcaatgcac
     4501 caggtcatca ttgacccagg cgtgttccac caggccgctg cctcgcaact cttcgcaggc
     4561 ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc acatgaggcg
     4621 gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt cgaacatccg
     4681 tcgggccgtc ggcgacagct tgcggtactt ctcccatatg aatttcgtgt agtggtcgcc
     4741 agcaaacagc acgacgattt cctcgtcgat caggacctgg caacgggacg ttttcttgcc
     4801 acggtccagg acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc caacgcggtc
     4861 ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg ccttcgtgta
     4921 ataccggcca ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg tgaaggtgat
     4981 cggctcgccg ataggggtgc gcttcgcgta ctccaacacc tgctgccaca ccagttcgtc
     5041 atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc acgtccttgt tgacgtggaa
     5101 aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg tgaacagggc
     5161 agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg caatatcgaa
     5221 caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg cggcctgctt
     5281 ggcctcgctg acctgttttg ccaggtcctc gccggcggtt tttcgcttct tggtcgtcat
     5341 agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct gttcgagacg
     5401 acgcgaacgc tccacggcgg ccgatggcgc gggcagggca gggggagcca gttgcacgct
     5461 gtcgcgctcg atcttggccg tagcttgctg gaccatcgag ccgacggact ggaaggtttc
     5521 gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg cggaaaaccc
     5581 cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac gtccgattca ttcaccctcc
     5641 ttgcgggatt gccccgactc acgccggggc aatgtgccct tattcctgat ttgacccgcc
     5701 tggtgccttg gtgtccagat aatccacctt atcggcaatg aagtcggtcc cgtagaccgt
     5761 ctggccgtcc ttctcgtact tggtattccg aatcttgccc tgcacgaata ccagctccgc
     5821 gaagtcgctc ttcttgatgg agcgcatggg gacgtgcttg gcaatcacgc gcaccccccg
     5881 gccgttttag cggctaaaaa agtcatggct ctgccctcgg gcggaccacg cccatcatga
     5941 ccttgccaag ctcgtcctgc ttctcttcga tcttcgccag cagggcgagg atcgtggcat
     6001 caccgaaccg cgccgtgcgc gggtcgtcgg tgagccagag tttcagcagg ccgcccaggc
     6061 ggcccaggtc gccattgatg cgggccagct cgcggacgtg ctcatagtcc acgacgcccg
     6121 tgattttgta gccctggccg acggccagca ggtaggccta caggctcatg ccggccgccg
     6181 ccgccttttc ctcaatcgct cttcgttcgt ctggaaggca gtacaccttg ataggtgggc
     6241 tgcccttcct ggttggcttg gtttcatcag ccatccgctt gccctcatct gttacgccgg
     6301 cggtagccgg ccagcctcgc agagcaggat tcccgttgag caccgccagg tgcgaataag
     6361 ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct
     6421 gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc
     6481 gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat gaccccgaag cagggttatg
     6541 cagcggaaaa gatccgtc
//
