LOCUS       pEML41003               5430 bp ds-DNA     circular     16-AUG-2023
DEFINITION  .
FEATURES             Location/Qualifiers
     protein_bind    79..100
                     /label="CAP binding site"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
                     /note="bound_moiety: E. coli catabolite activator protein note: CAP binding activates transcription in the presence of cAMP."
     promoter        115..145
                     /label="lac promoter"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /note="note: promoter for the E. coli lac operon"
     protein_bind    153..169
                     /label="lac operator"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="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)."
     primer_bind     158..180
                     /label="M13/pUC Reverse"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="note: In lacZ gene"
     primer_bind     177..193
                     /label="M13 rev"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="note: common sequencing primer, one of multiple similar variants"
     primer_bind     177..193
                     /label="M13 Reverse"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="note: In lacZ gene. Also called M13-rev"
     CDS             189..512
                     /label="lacZ-alpha"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="gene: lacZ fragment product: LacZ-alpha fragment of beta-galactosidase translation: MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDAA"
     misc_feature    206..262
                     /label="MCS"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: pUC18/19 multiple cloning site"
     primer_bind     complement(263..279)
                     /label="M13 fwd"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="note: common sequencing primer, one of multiple similar variants"
     primer_bind     complement(488..507)
                     /label="pRS-marker"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: pRS vectors, use to sequence yeast selectable marker"
     terminator      617..711
                     /label="lambda t0 terminator"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /note="note: transcription terminator from phage lambda"
     primer          complement(629..650)
                     /label="BEVA Lv1 REV"
                     /note="sequence: tctgaggtcattactggatcta"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer_bind     complement(810..829)
                     /label="pENTR-R"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /note="note: pENTR vectors, reverse primer"
     CDS             821..1636
                     /label="KanR"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="product: aminoglycoside phosphotransferase note: confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes translation: MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    1652..3197
                     /label="Position 3 module for suicide plasmid"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     rep_origin      complement(1950..2495)
                     /label="p15A ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="direction: LEFT note: Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin."
     misc_feature    complement(2309..2613)
                     /label="TRAJ"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     oriT            2869..2978
                     /label="oriT"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: incP origin of transfer"
     misc_feature    3202..3353
                     /label="Position 3 module for suicide plasmid"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     promoter        3358..3803
                     /label="sacB promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="gene: sacR note: sacB promoter and control region"
     misc_feature    3358..5317
                     /label="Position 4 module for suicide plasmid"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             3804..3890
                     /label="SacB"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
                     /note="codon_start: 1 gene: Bacillus subtilis sacB note: negative selection marker product: secreted levansucrase that renders bacterial growth sensitive to sucrose protein_id: transl_table: 1 translation: MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYRGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGISSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"
     CDS             3891..5225
                     /label="SacB"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
                     /note="codon_start: 1 gene: Bacillus subtilis sacB note: negative selection marker product: secreted levansucrase that renders bacterial growth sensitive to sucrose protein_id: transl_table: 1 translation: MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYRGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGISSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"
     primer          5305..5325
                     /label="pEML41003-seq-F"
                     /note="sequence: cttgaacgaattgCAGAtcta"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     terminator      5335..5421
                     /label="rrnB T1 terminator"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /note="gene: Escherichia coli rrnB note: transcription terminator T1 from the E. coli rrnB gene"
     primer          5421..7
                     /label="BEVA-Lv1-seq-F-v3"
                     /note="sequence: gatgcctgccgaattcg"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
ORIGIN
        1 gaattcggat ccggagtgag accgcagctg gcacgacagg tttgccgact ggaaagcggg
       61 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      121 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      181 aaacagctat gaccatgatt acgccaagct tgcatgcctg caggtcgact ctagaggatc
      241 cccgggtacc gagctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
      301 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
      361 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
      421 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
      481 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
      541 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
      601 cggtctcacg ctcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
      661 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaggcaat
      721 tgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
      781 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcagcgtga
      841 aacgagctgt agccgtccgc gtctgaacag caacatggat gcggatctgt atggctataa
      901 atgggcgcgt gataacgtgg gtcagagcgg cgcgaccatt tatcgtctgt atggcaaacc
      961 ggatgcgccg gaactgtttc tgaaacatgg caaaggcagc gtggcgaacg atgtgaccga
     1021 tgaaatggtg cgtctgaact ggctgaccga atttatgccg ctgccgacca ttaaacattt
     1081 tattcgcacc ccggatgatg cgtggctgct gaccaccgcg attccgggca aaaccgcgtt
     1141 tcaggtgctg gaagaatatc cggatagcgg cgaaaacatt gtggatgcgc tggccgtgtt
     1201 tctgcgtcgt ctgcatagca ttccggtgtg caactgcccg tttaacagcg atcgtgtgtt
     1261 tcgtctggcc caggcgcaga gccgtatgaa caacggcctg gtggatgcga gcgattttga
     1321 tgatgaacgt aacggctggc cggtggaaca ggtgtggaaa gaaatgcata aactgctgcc
     1381 gtttagcccg gatagcgtgg tgacccacgg cgattttagc ctggataacc tgattttcga
     1441 tgaaggcaaa ctgattggct gcattgatgt gggccgtgtg ggcattgcgg atcgttatca
     1501 ggatctggcc attctgtgga actgcctggg cgaatttagc ccgagcctgc aaaaacgtct
     1561 gtttcagaaa tatggcattg ataatccgga tatgaacaaa ctgcaatttc atctgatgct
     1621 ggatgaattt ttctaataat taattgaact atgcgtaacg gcaaaagcac cgccggacat
     1681 cagcgctagc ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag
     1741 tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag
     1801 gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
     1861 ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag
     1921 ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat
     1981 cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag
     2041 gcgtttCccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt
     2101 gtcattccgc tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta
     2161 ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtCcg accgctgcgc
     2221 cttatccggt aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc
     2281 agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct
     2341 aaactgaaag gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag
     2401 agttggtagc tcagagaacc ttcgAaaaac cgccctgcaa ggcggttttt tcgttttcag
     2461 agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aatcagataa
     2521 aatatttcta gctagagtcg atcttcgcca gcagggcGag gatcgtggca tcaccgaacc
     2581 gcgccgtGCg cGgGtcgtcg gtgagccaga gtttcagcag gccgcccagg tcgccattga
     2641 tgcgggccag ctcgcggacg tgctcatagt ccacgacgcc cgtgattttg tagccctggc
     2701 cgacgGCcAG caggtaggcc gacaGgctca tgccggccgc cgccgccttt tcctcaatcg
     2761 ctcttcgttc gtctggaagg cagtacacct tgataggtgg gctgcccttc ctggttggct
     2821 tggtttcatc agccatccgc ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc
     2881 gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac
     2941 acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc
     3001 aaggaaagtc tacAcgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat
     3061 ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa aagcgctgct
     3121 tccctgctgt tttgtggaat atctaccgac tggaaacagg caaatgcagg aaattactga
     3181 actgagggga caggcgaGAC Acgatgccaa agagctacac cgacgagctg gccgagtggg
     3241 ttgaatcccg cgcggccaag aagcgccggc gtgatgaggc tgcggttgcg ttcctggcgg
     3301 tgagggcgga tgtcgatcga ctctagctag aggatcgatc ctttttaacc catTTACcac
     3361 atatacctgc cgttcactat tatttagtga aatgagatat tatgatattt tctgaattgt
     3421 gattaaaaag gcaactttat gcccatgcaa cagaaactat aaaaaataca gagaatgaaa
     3481 agaaacagat agatttttta gttctttagg cccgtagtct gcaaatcctt ttatgatttt
     3541 ctatcaaaca aaagaggaaa atagaccagt tgcaatccaa acgagagtct aatagaatga
     3601 ggtcgaaaag taaatcgcgc gggtttgtta ctgataaagc aggcaagacc taaaatgtgt
     3661 aaagggcaaa gtgtatactt tggcgtcacc ccttacatat tttaggtctt tttttattgt
     3721 gcgtaactaa cttgccatct tcaaacagga gggctggaag aagcagaccg ctaacacagt
     3781 acataaaaaa ggagacatga acgatgaaca tcaaaaagtt tgcaaaacaa gcaacagtat
     3841 taacctttac taccgcactg ctggcaggag gcgcaactca agcgtttgcg aaagaaacga
     3901 accaaaagcc atataaggaa acatacggca tttcccatat tacacgccat gatatgctgc
     3961 aaatccctga acagcaaaaa aatgaaaaat atcaagttcc tgaattcgat tcgtccacaa
     4021 ttaaaaatat ctcttctgca aaaggcctgg acgtttggga cagctggcca ttacaaaacg
     4081 ctgacggcac tgtcgcaaac tatcacggct accacatcgt ctttgcatta gccggagatc
     4141 ctaaaaatgc ggatgacaca tcgatttaca tgttctatca aaaagtcggc gaaacttcta
     4201 ttgacagctg gaaaaacgct ggccgcgtct ttaaagacag cgacaaattc gatgcaaatg
     4261 attctatcct aaaagaccaa acacaagaat ggtcaggttc agccacattt acatctgacg
     4321 gaaaaatccg tttattctac actgatttct ccggtaaaca ttacggcaaa caaacactga
     4381 caactgcaca agttaacgta tcagcatcag acagctcttt gaacatcaac ggtgtagagg
     4441 attataaatc aatctttgac ggtgacggaa aaacgtatca aaatgtacag cagttcatcg
     4501 atgaaggcaa ctacagctca ggcgacaacc atacgctgag agatcctcac tacgtagaag
     4561 ataaaggcca caaatactta gtatttgaag caaacactgg aactgaagat ggctaccaag
     4621 gcgaagaatc tttatttaac aaagcatact atggcaaaag cacatcattc ttccgtcaag
     4681 aaagtcaaaa acttctgcaa agcgataaaa aacgcacggc tgagttagca aacggcgctc
     4741 tcggtatgat tgagctaaac gatgattaca cactgaaaaa agtgatgaaa ccgctgattg
     4801 catctaacac agtaacagat gaaattgaac gcgcgaacgt ctttaaaatg aacggcaaat
     4861 ggtacctgtt cactgactcc cgcggatcaa aaatgacgat tgacggcatt acgtctaacg
     4921 atatttacat gcttggttat gtttctaatt ctttaactgg cccatacaag ccgctgaaca
     4981 aaactggcct tgtgttaaaa atggatcttg atcctaacga tgtaaccttt acttactcac
     5041 acttcgctgt acctcaagcg aaaggaaaca atgtcgtgat tacaagctat atgacaaaca
     5101 gaggattcta cgcagacaaa caatcaacgt ttgcgccaag cttcctgctg aacatcaaag
     5161 gcaagaaaac atctgttgtc aaagacagca tccttgaaca aggacaatta acagttaaca
     5221 aataaaaacg caaaagaaaa tgccgatggc cgcggcgttg tgacaattta ccgaacaact
     5281 ccgcggccgg gaagccgatc tcggcttgaa cgaattgCAG Atctagggcg gcggatttgt
     5341 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga
     5401 ctgagccttt cgttttattt gatgcctgcc
//