LOCUS       pG1AK-sfGFP                5650 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Modular shuttle vector for Geobacillus and E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    pG1AK-sfGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5650)
  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 5650)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..5650
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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(62..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             complement(1198..2058)
                     /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     1821..1840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2059..2163)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2231..2249
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             4874..5587
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
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 atggccggcc ctgacagtta
     1201 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1261 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1321 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1381 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1441 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1501 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1561 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1621 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1681 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1741 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1801 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1861 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1921 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1981 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2041 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2101 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2161 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     2221 aacctataaa aataggcgta tcacgaggcc gcggccgcct gtcgagaatt tttggaaaat
     2281 agaccttgtc gcgactcgaa gattagtagt acaatagaga caacataaca gaacgggaca
     2341 aaatagaaga gcttccgaag cgtgtgtaag tggtgctccc aacaccgttc gccgtactcg
     2401 ccctacggca aacacctgct cgaaagctct tgtgttaaca tatttatgcc ttgtcttcat
     2461 tttaatgcat agatatgttg aatgcaagag cgaaacgctt ggaaattttc gtttttcctc
     2521 cgatgagagc gtgtttgcct gcgtattaat gtacggcaac gcgctctttt gtttgtcccg
     2581 aaaaattcgg gagaaacggg aggagaacgg aaatgtacat aatcactagc gaacaattca
     2641 cgcaactgaa agaaatcgag agagtcacag atttcgttta taaacgccgt aagattaaaa
     2701 atgagctgat ggagcgtatc gccgacgcat taggccgcaa gttccgcaaa gcgcaaaaaa
     2761 caacgcgcgt catcgacgaa attgtttggc tcgcaacaga ccgcggtttt tcgttcccag
     2821 ggcgcgaaac gcttgccaaa aagttaggcg tgtccttgcc gactgttgat cgtgctactc
     2881 gcatgctcaa agactcgggc gaagtcgtcg tttgctatcg tgagaatccg aatagtaacg
     2941 gacctaaaac gcctgtattc attttccgga gccatgcaaa tttcgagcgt attgcagccg
     3001 ttttaaactt gcgtgacaac gaagctgata aagtagaaaa cgccgaaaag cctacggaat
     3061 caagcgattt agcgcgtaaa acggacgcta ccattagttc accagttgaa aaacatgacg
     3121 atattaataa attaatagac aatacacatc cgttggataa aattgtaaaa tacgtcacta
     3181 ttaaagtgaa cgaagtgcag aagcgtcatc cttacggcat tcgatatcta tccgcataca
     3241 taaacaaaac actcgacgac ctaatgcaac gtgcaaaagc gaaggtcaaa gccgaaaaag
     3301 ctcgcgccca acaacgcaaa caagccgact atcaaccgac cgcgaccggc ttaatttggt
     3361 acgacttcac ggcggacaca agcgccagcc agccgacaag ccagcgcaag ccagcgtttg
     3421 acagcgaaga catgtttgcg ccgttattgc aggctgttgc acgaattaag ggcgttgagg
     3481 tgtgacacac tcgttttaag gcgtctaaca ggcgcgtaga gacgtcttta agtcggggga
     3541 atattaaaaa ttccacgcct cctaaaacgc gatatacgcc gattatgagt ttaaacgaag
     3601 attagatgct ataattgtta ttaaaaggat tgaaggatgc ttaggaagac gagttattaa
     3661 tagctgaata agaacggtgc tctccaaata ttcttattta gaaaagcaaa tctaaaatta
     3721 tctgaaaagg gaatgagaat agtgaatgga ccaataataa tgactagaga agaaagaatg
     3781 aagattgttc atgaaattaa ggaacgaata ttggataaat atggggatga tgttaaggct
     3841 attggtgttt atggctctct tggtcgtcag actgatgggc cctattcgga tattgagatg
     3901 atgtgtgtca tgtcaacaga ggaagcagag ttcagccatg aatggacaac cggtgagtgg
     3961 aaggtggaag tgaattttta tagcgaagag attctactag attatgcatc tcaggtggaa
     4021 tcagattggc cgcttacaca tggtcaattt ttctctattt tgccgattta tgattcaggt
     4081 ggatacttag agaaagtgta tcaaactgct aaatcggtag aagcccaaaa gttccacgat
     4141 gcgatttgtg cccttatcgt agaagagctg tttgaatatg caggcaaatg gcgtaatatt
     4201 cgtgtgcaag gaccgacaac atttctacca tccttgactg tacaggtagc aatggcaggt
     4261 gccatgttga ttggtctgca tcatcgcatc tgttatacga cgagcgcttc ggtcttaact
     4321 gaagcagtta agcaatcaga tcttccttca ggttatgacc atctgtgcca gttcgtaatg
     4381 tctggtcaac tttccgactc tgagaaactt ctggaatcgc tagagaattt ctggaatggg
     4441 attcaggagt ggacagaacg acacggatat atagtggatg tgtcaaaacg cataccattt
     4501 tgataaggcg cgcctctacg gaccgagtat tcaacaaacg ggccagtttg ttgaaatcgt
     4561 taattaagcg tactagtcct aggctagctg ataccaggtc agtcccggga caggtcgacc
     4621 tcgagtacgc ctgcaggaac aatcgttaaa gcggacgttt ttgcgccgcc cggatttgct
     4681 tgaaaactac ccgctgacag aaaagcaaaa acgatggatc gaagagtgga aaaaagaaaa
     4741 acagtagcta ttgcgcatga tacaagttta tgctactata ttccttgtgc aacttaagcg
     4801 atttgcttaa gcgaggaaaa cgatgttccg ctgcaatgat gaaaaagcat tgtctagata
     4861 aggagtgatt cgaatgcgta aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt
     4921 ggaactggat ggtgatgtca acggtcataa gttttccgtg cgtggcgagg gtgaaggtga
     4981 cgcaactaat ggtaaactga cgctgaagtt catctgtact actggtaaac tgccggtacc
     5041 ttggccgact ctggtaacga cgctgactta tggtgttcag tgctttgctc gttatccgga
     5101 ccatatgaag cagcatgact tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg
     5161 cacgatttcc tttaaggatg acggcacgta caaaacgcgt gcggaagtga aatttgaagg
     5221 cgataccctg gtaaaccgca ttgagctgaa aggcattgac tttaaagaag acggcaatat
     5281 cctgggccat aagctggaat acaattttaa cagccacaat gtttacatca ccgccgataa
     5341 acaaaaaaat ggcattaaag cgaattttaa aattcgccac aacgtggagg atggcagcgt
     5401 gcagctggct gatcactacc agcaaaacac tccaatcggt gatggtcctg ttctgctgcc
     5461 agacaatcac tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga
     5521 tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc acgcatggta tggatgaact
     5581 gtacaaatga taagagctca atttcgcgcc ccgaaagggg cgtttttttg cgaagcctaa
     5641 ggtatcattt
//