LOCUS       sequence-135061-        5848 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 5848)
  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 5848)
  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-PheB
FEATURES             Location/Qualifiers
     source          1..5848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(71..87)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(71..87)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(84..106)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    95..111
                     /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(119..125,126..143,144..149))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    164..185
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(302..319)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(473..1061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(553..572)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1255..2046,2047..2115))
                     /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     1878..1897
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2116..2220)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2288..2306
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gagctcaatt tcgcgccccg aaaggggcgt ttttttgcga agcctaaggt atcatttaaa
       61 tgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
      121 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      181 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      241 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
      301 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      361 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      421 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      481 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      541 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      601 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      661 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
      721 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
      781 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
      841 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
      901 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
      961 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1021 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1081 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1141 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1201 aaagtatata tgagtaaact tggtctgaca gttaccaatg gccggccctg acagttacca
     1261 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1321 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1381 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1441 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1501 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1561 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1621 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1681 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1741 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1801 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1861 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1921 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1981 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     2041 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2101 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2161 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2221 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     2281 ctataaaaat aggcgtatca cgaggccgcg gccgcctgtc gagaattttt ggaaaataga
     2341 ccttgtcgcg actcgaagat tagtagtaca atagagacaa cataacagaa cgggacaaaa
     2401 tagaagagct tccgaagcgt gtgtaagtgg tgctcccaac accgttcgcc gtactcgccc
     2461 tacggcaaac acctgctcga aagctcttgt gttaacatat ttatgccttg tcttcatttt
     2521 aatgcataga tatgttgaat gcaagagcga aacgcttgga aattttcgtt tttcctccga
     2581 tgagagcgtg tttgcctgcg tattaatgta cggcaacgcg ctcttttgtt tgtcccgaaa
     2641 aattcgggag aaacgggagg agaacggaaa tgtacataat cactagcgaa caattcacgc
     2701 aactgaaaga aatcgagaga gtcacagatt tcgtttataa acgccgtaag attaaaaatg
     2761 agctgatgga gcgtatcgcc gacgcattag gccgcaagtt ccgcaaagcg caaaaaacaa
     2821 cgcgcgtcat cgacgaaatt gtttggctcg caacagaccg cggtttttcg ttcccagggc
     2881 gcgaaacgct tgccaaaaag ttaggcgtgt ccttgccgac tgttgatcgt gctactcgca
     2941 tgctcaaaga ctcgggcgaa gtcgtcgttt gctatcgtga gaatccgaat agtaacggac
     3001 ctaaaacgcc tgtattcatt ttccggagcc atgcaaattt cgagcgtatt gcagccgttt
     3061 taaacttgcg tgacaacgaa gctgataaag tagaaaacgc cgaaaagcct acggaatcaa
     3121 gcgatttagc gcgtaaaacg gacgctacca ttagttcacc agttgaaaaa catgacgata
     3181 ttaataaatt aatagacaat acacatccgt tggataaaat tgtaaaatac gtcactatta
     3241 aagtgaacga agtgcagaag cgtcatcctt acggcattcg atatctatcc gcatacataa
     3301 acaaaacact cgacgaccta atgcaacgtg caaaagcgaa ggtcaaagcc gaaaaagctc
     3361 gcgcccaaca acgcaaacaa gccgactatc aaccgaccgc gaccggctta atttggtacg
     3421 acttcacggc ggacacaagc gccagccagc cgacaagcca gcgcaagcca gcgtttgaca
     3481 gcgaagacat gtttgcgccg ttattgcagg ctgttgcacg aattaagggc gttgaggtgt
     3541 gacacactcg ttttaaggcg tctaacaggc gcgtagagac gtctttaagt cgggggaata
     3601 ttaaaaattc cacgcctcct aaaacgcgat atacgccgat tatgagttta aacgaagatt
     3661 agatgctata attgttatta aaaggattga aggatgctta ggaagacgag ttattaatag
     3721 ctgaataaga acggtgctct ccaaatattc ttatttagaa aagcaaatct aaaattatct
     3781 gaaaagggaa tgagaatagt gaatggacca ataataatga ctagagaaga aagaatgaag
     3841 attgttcatg aaattaagga acgaatattg gataaatatg gggatgatgt taaggctatt
     3901 ggtgtttatg gctctcttgg tcgtcagact gatgggccct attcggatat tgagatgatg
     3961 tgtgtcatgt caacagagga agcagagttc agccatgaat ggacaaccgg tgagtggaag
     4021 gtggaagtga atttttatag cgaagagatt ctactagatt atgcatctca ggtggaatca
     4081 gattggccgc ttacacatgg tcaatttttc tctattttgc cgatttatga ttcaggtgga
     4141 tacttagaga aagtgtatca aactgctaaa tcggtagaag cccaaaagtt ccacgatgcg
     4201 atttgtgccc ttatcgtaga agagctgttt gaatatgcag gcaaatggcg taatattcgt
     4261 gtgcaaggac cgacaacatt tctaccatcc ttgactgtac aggtagcaat ggcaggtgcc
     4321 atgttgattg gtctgcatca tcgcatctgt tatacgacga gcgcttcggt cttaactgaa
     4381 gcagttaagc aatcagatct tccttcaggt tatgaccatc tgtgccagtt cgtaatgtct
     4441 ggtcaacttt ccgactctga gaaacttctg gaatcgctag agaatttctg gaatgggatt
     4501 caggagtgga cagaacgaca cggatatata gtggatgtgt caaaacgcat accattttga
     4561 taaggcgcgc ctctacggac cgagtattca acaaacgggc cagtttgttg aaatcgttaa
     4621 ttaagcgtac tagtcctagg ctagctgata ccaggtcagt cccgggacag gtcgacctcg
     4681 agtacgcctg caggaacaat cgttaaagcg gacgtttttg cgccgcccgg atttgcttga
     4741 aaactacccg ctgacagaaa agcaaaaacg atggatcgaa gagtggaaaa aagaaaaaca
     4801 gtagctattg cgcatgatac aagtttatgc tactatattc cttgtgcaac ttaagcgatt
     4861 tgcttaagcg aggaaaacga tgttccgctg caatgatgaa aaagcattgt ctagataagg
     4921 agtgattcga atggctatta tgcggatcgg caaggccgaa ataagagtca tggatctcga
     4981 agaatctgtc aagtattaca cgaatgtgat tggcctggag gaagtgggaa ggagtgaagg
     5041 aagagtttat ttaaaggcat gggatgaatt cgatcaccat agcctcattc ttcaagaagc
     5101 cgattcccct ggccttgatc acattgcttt taaggtggaa catgaagacg atttagccaa
     5161 gtacgagaag aagatcgagc aattcgggtg tacgttaaaa cggatttcca aagggacaag
     5221 gcttgcagaa ggagaagcaa ttcgcttcga gcttccaaca gggcatcaag tggaattgta
     5281 ccatgaaatc gtgcgtgtag gcacgaagac aggaaatttg aatccagccc catggccgga
     5341 tggaatgcgc gggattgcac cgcaccgcct agatcactta gcgctgacag gagaagatat
     5401 caacacagtg acaagatttt tcacagaagc cttggatatg ttcattagcg aaaaaattat
     5461 gacagtagat ggggaagaga tggtagggag ctttatattt gccagaaacg gaaaagcgca
     5521 cgatgttgcc tttattaaag ggccagataa gaaaatgcat catgtcgcat tctatgtgga
     5581 caattggtat gaagtgttaa aggcagcgga tattttatcc aaaaataatg tccaattcga
     5641 tgtgacaccg acccgccatg ggattacgcg tggacaaacc acctacttct ttgatccttc
     5701 aggtaatcgc aatgaagctt ttgcaagcgg ttacattacg tatcctgatt ttcctaccat
     5761 aacatggaca gaagacaaaa tcggtcaagg aatcttctat catagaagag aattgacgga
     5821 gtcattcatc aaggcgctga cataataa
//
