LOCUS       sequence-130147-        3841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modular shuttle vector for Geobacillus and E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3841)
  AUTHORS   Reeve B, Martinez-Klimova E, de Jonghe J, Leak DJ, Ellis T
  TITLE     The Geobacillus Plasmid Set: A Modular Toolkit for Thermophile 
            Engineering.
  JOURNAL   ACS Synth Biol. 2016 Jul 1.
  PUBMED    27332993
REFERENCE   2  (bases 1 to 3841)
  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. 2016 Jul 1."
COMMENT     Sequence Label: pG2K
FEATURES             Location/Qualifiers
     source          1..3841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(14..30)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14..30)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(27..49)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    38..54
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(62..68,69..86,87..92))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(245..262)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(416..1004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(496..515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1662..2666
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
ORIGIN
        1 aaatgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
       61 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
      121 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
      181 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
      241 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      301 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
      361 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      421 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      481 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      541 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      601 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      661 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      721 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      781 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      841 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      901 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
      961 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1021 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1081 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1141 tctaaagtat atatgagtaa acttggtctg acagttacca atgcggccgc ctaaaataag
     1201 cgatttaaag cgtgatgtga gtttaatcta tgaaagcact aaggaattcc ttaaggaacg
     1261 tacagacggc ttaaaagcct ttaaaaacgt ttttaagggg tttgtagaca aggtaaagga
     1321 taaaacagca caattccaag aaaaacacga tttagaacct aaaaagaacg aatttgaact
     1381 aactcataac cgagaggtaa aaaaagaacg aagtcgagat cagggaatga gtttataaaa
     1441 taaaaaaagc acctgaaaag gtgtcttttt ttgatggttt tgaacttgtt ctttcttatc
     1501 ttgatacata tagaaataac gtcattttta ttttagttgc tgaaaggtgc gttgaagtgt
     1561 tggtatgtat gtgttttaaa gtattgaaaa cccttaaaat tggttgcaca gaaaaacccc
     1621 atctgttaaa gttataagtg actaaacaaa taactaaata gatgggggtt tcttttaata
     1681 ttatgtgtcc taatagtagc atttattcag atgaaaaatc aagggtttta gtggacaaga
     1741 caaaaagtgg aaaagtgaga ccatggagag aaaagaaaat cgctaatgtt gattactttg
     1801 aacttctgca tattcttgaa tttaaaaagg ctgaaagagt aaaagattgt gctgaaatat
     1861 tagagtataa acaaaatcgt gaaacaggcg aaagaaagtt gtatcgagtg tggttttgta
     1921 aatccaggct ttgtccaatg tgcaactgga ggagagcaat gaaacatggc attcagtcac
     1981 aaaaggttgt tgctgaagtt attaaacaaa agccaacagt tcgttggttg tttctcacat
     2041 taacagttaa aaatgtttat gatggcgaag aattaaataa gagtttgtca gatatggctc
     2101 aaggatttcg ccgaatgatg caatataaaa aaattaataa aaatcttgtt ggttttatgc
     2161 gtgcaacgga agtgacaata aataataaag ataattctta taatcagcac atgcatgtat
     2221 tggtatgtgt ggaaccaact tattttaaga atacagaaaa ctacgtgaat caaaaacaat
     2281 ggattcaatt ttggaaaaag gcaatgaaat tagactatga tccaaatgta aaagttcaaa
     2341 tgattcgacc gaaaaataaa tataaatcgg atatacaatc ggcaattgac gaaactgcaa
     2401 aatatcctgt aaaggatacg gattttatga ccgatgatga agaaaagaat ttgaaacgtt
     2461 tgtctgattt ggaggaaggt ttacaccgta aaaggttaat ctcctatggt ggtttgttaa
     2521 aagaaataca taaaaaatta aaccttgatg acacagaaga aggcgatttg attcatacag
     2581 atgatgacga aaaagccgat gaagatggat tttctattat tgcaatgtgg aattgggaac
     2641 ggaaaaatta ttttattaaa gagtagttca acaaacgggc cagtttgttg aagattagat
     2701 gctataattg gtttaaacga agattagatg ctataattgt tattaaaagg attgaaggat
     2761 gcttaggaag acgagttatt aatagctgaa taagaacggt gctctccaaa tattcttatt
     2821 tagaaaagca aatctaaaat tatctgaaaa gggaatgaga atagtgaatg gaccaataat
     2881 aatgactaga gaagaaagaa tgaagattgt tcatgaaatt aaggaacgaa tattggataa
     2941 atatggggat gatgttaagg ctattggtgt ttatggctct cttggtcgtc agactgatgg
     3001 gccctattcg gatattgaga tgatgtgtgt catgtcaaca gaggaagcag agttcagcca
     3061 tgaatggaca accggtgagt ggaaggtgga agtgaatttt tatagcgaag agattctact
     3121 agattatgca tctcaggtgg aatcagattg gccgcttaca catggtcaat ttttctctat
     3181 tttgccgatt tatgattcag gtggatactt agagaaagtg tatcaaactg ctaaatcggt
     3241 agaagcccaa aagttccacg atgcgatttg tgcccttatc gtagaagagc tgtttgaata
     3301 tgcaggcaaa tggcgtaata ttcgtgtgca aggaccgaca acatttctac catccttgac
     3361 tgtacaggta gcaatggcag gtgccatgtt gattggtctg catcatcgca tctgttatac
     3421 gacgagcgct tcggtcttaa ctgaagcagt taagcaatca gatcttcctt caggttatga
     3481 ccatctgtgc cagttcgtaa tgtctggtca actttccgac tctgagaaac ttctggaatc
     3541 gctagagaat ttctggaatg ggattcagga gtggacagaa cgacacggat atatagtgga
     3601 tgtgtcaaaa cgcataccat tttgataagg cgcgcctcta cggaccgagt attcaacaaa
     3661 cgggccagtt tgttgaaatc gttaattaag cgtactagtc ctaggctagc tgataccagg
     3721 tcagtcccgg gacaggtcga cctcgagtac gcctgcagga gtgttctaga ctagaagctt
     3781 cactgagctc aatttcgcgc cccgaaaggg gcgttttttt gcgaagccta aggtatcatt
     3841 t
//
