LOCUS       sequence-130104-        4719 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 4719)
  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 4719)
  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: pG1AK
FEATURES             Location/Qualifiers
     source          1..4719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(225..241)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(225..241)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(238..260)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    249..265
                     /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(273..279,280..297,298..303))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    318..339
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(456..473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(627..1215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(707..726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1409..2200,2201..2269))
                     /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     2032..2051
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2270..2374)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2442..2460
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgcgcctcta cggaccgagt attcaacaaa cgggccagtt tgttgaaatc gttaattaag
       61 cgtactagtc ctaggctagc tgataccagg tcagtcccgg gacaggtcga cctcgagtac
      121 gcctgcagga gtgttctaga ctagaagctt cactgagctc aatttcgcgc cccgaaaggg
      181 gcgttttttt gcgaagccta aggtatcatt taaatgtaat catggtcata gctgtttcct
      241 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
      301 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
      361 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
      421 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
      481 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
      541 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
      601 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
      661 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
      721 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
      781 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
      841 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
      901 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
      961 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1021 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1081 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1141 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1201 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1261 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1321 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     1381 gacagttacc aatggccggc cctgacagtt accaatgctt aatcagtgag gcacctatct
     1441 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1501 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1561 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1621 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1681 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1741 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1801 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1861 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1921 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1981 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2041 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2101 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2161 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2221 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2281 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2341 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2401 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     2461 cgcggccgcc tgtcgagaat ttttggaaaa tagaccttgt cgcgactcga agattagtag
     2521 tacaatagag acaacataac agaacgggac aaaatagaag agcttccgaa gcgtgtgtaa
     2581 gtggtgctcc caacaccgtt cgccgtactc gccctacggc aaacacctgc tcgaaagctc
     2641 ttgtgttaac atatttatgc cttgtcttca ttttaatgca tagatatgtt gaatgcaaga
     2701 gcgaaacgct tggaaatttt cgtttttcct ccgatgagag cgtgtttgcc tgcgtattaa
     2761 tgtacggcaa cgcgctcttt tgtttgtccc gaaaaattcg ggagaaacgg gaggagaacg
     2821 gaaatgtaca taatcactag cgaacaattc acgcaactga aagaaatcga gagagtcaca
     2881 gatttcgttt ataaacgccg taagattaaa aatgagctga tggagcgtat cgccgacgca
     2941 ttaggccgca agttccgcaa agcgcaaaaa acaacgcgcg tcatcgacga aattgtttgg
     3001 ctcgcaacag accgcggttt ttcgttccca gggcgcgaaa cgcttgccaa aaagttaggc
     3061 gtgtccttgc cgactgttga tcgtgctact cgcatgctca aagactcggg cgaagtcgtc
     3121 gtttgctatc gtgagaatcc gaatagtaac ggacctaaaa cgcctgtatt cattttccgg
     3181 agccatgcaa atttcgagcg tattgcagcc gttttaaact tgcgtgacaa cgaagctgat
     3241 aaagtagaaa acgccgaaaa gcctacggaa tcaagcgatt tagcgcgtaa aacggacgct
     3301 accattagtt caccagttga aaaacatgac gatattaata aattaataga caatacacat
     3361 ccgttggata aaattgtaaa atacgtcact attaaagtga acgaagtgca gaagcgtcat
     3421 ccttacggca ttcgatatct atccgcatac ataaacaaaa cactcgacga cctaatgcaa
     3481 cgtgcaaaag cgaaggtcaa agccgaaaaa gctcgcgccc aacaacgcaa acaagccgac
     3541 tatcaaccga ccgcgaccgg cttaatttgg tacgacttca cggcggacac aagcgccagc
     3601 cagccgacaa gccagcgcaa gccagcgttt gacagcgaag acatgtttgc gccgttattg
     3661 caggctgttg cacgaattaa gggcgttgag gtgtgacaca ctcgttttaa ggcgtctaac
     3721 aggcgcgtag agacgtcttt aagtcggggg aatattaaaa attccacgcc tcctaaaacg
     3781 cgatatacgc cgattatgag tttaaacgaa gattagatgc tataattgtt attaaaagga
     3841 ttgaaggatg cttaggaaga cgagttatta atagctgaat aagaacggtg ctctccaaat
     3901 attcttattt agaaaagcaa atctaaaatt atctgaaaag ggaatgagaa tagtgaatgg
     3961 accaataata atgactagag aagaaagaat gaagattgtt catgaaatta aggaacgaat
     4021 attggataaa tatggggatg atgttaaggc tattggtgtt tatggctctc ttggtcgtca
     4081 gactgatggg ccctattcgg atattgagat gatgtgtgtc atgtcaacag aggaagcaga
     4141 gttcagccat gaatggacaa ccggtgagtg gaaggtggaa gtgaattttt atagcgaaga
     4201 gattctacta gattatgcat ctcaggtgga atcagattgg ccgcttacac atggtcaatt
     4261 tttctctatt ttgccgattt atgattcagg tggatactta gagaaagtgt atcaaactgc
     4321 taaatcggta gaagcccaaa agttccacga tgcgatttgt gcccttatcg tagaagagct
     4381 gtttgaatat gcaggcaaat ggcgtaatat tcgtgtgcaa ggaccgacaa catttctacc
     4441 atccttgact gtacaggtag caatggcagg tgccatgttg attggtctgc atcatcgcat
     4501 ctgttatacg acgagcgctt cggtcttaac tgaagcagtt aagcaatcag atcttccttc
     4561 aggttatgac catctgtgcc agttcgtaat gtctggtcaa ctttccgact ctgagaaact
     4621 tctggaatcg ctagagaatt tctggaatgg gattcaggag tggacagaac gacacggata
     4681 tatagtggat gtgtcaaaac gcataccatt ttgataagg
//
