LOCUS pX33 9681 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Allelic change mutagenesis.
ACCESSION .
VERSION .
KEYWORDS pX33
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9681)
AUTHORS Gebhard S, Tran SL, Cook GM
TITLE The Phn system of Mycobacterium smegmatis: a second high-affinity
ABC-transporter for phosphate.
JOURNAL Microbiology. 2006 Nov;152(Pt 11):3453-65.
PUBMED 17074913
REFERENCE 2 (bases 1 to 9681)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9681
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(3913..3929)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3913..3929)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(3961..3980)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3962..3980)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3990..4007)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3990..4006)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3999..4021)
/label=M13/pUC Forward
/note="In lacZ gene"
promoter 4545..4573
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
CDS 4762..5295
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
CDS complement(5388..6809)
/codon_start=1
/gene="Bacillus subtilis sacB"
/product="secreted levansucrase that renders bacterial
growth sensitive to sucrose"
/label=SacB
/note="negative selection marker"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
promoter complement(6810..7255)
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
oriT 7631..7740
/note="incP origin of transfer"
rep_origin 8123..8668
/direction=RIGHT
/label=p15A ori
/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.
"
primer_bind 8569..8588
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8785..8802
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 9061..9082
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9097..9127
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9135..9151
/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)."
primer_bind 9140..9162
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 9159..9175
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9159..9175
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 9194..9214
/label=T3
/note="T3 promoter, forward primer"
promoter 9196..9214
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind 9243..9259
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 9244..9260
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 atgaactatg aggaggatga cgtcatgaac aaaggtgtaa tgcgaccggg ccatgtgcag
61 ctgcgtgtac tggacatgag caaggccctg gaacactacg tcgagttgct gggcctgatc
121 gagatggacc gtgacgacca gggccgtgtc tatctgaagg cttggaccga agtggataag
181 ttttccctgg tgctacgcga ggctgacgag ccgggcatgg attttatggg tttcaaggtt
241 gtggatgagg atgctctccg gcaactggag cgggatctga tggcatatgg ctgtgccgtt
301 gagcagctac ccgcaggtga actgaacagt tgtggccggc gcgtgcgctt ccaggccccc
361 tccgggcatc acttcgagtt gtatgcagac aaggaatata ctggaaagtg gggtttgaat
421 gacgtcaatc ccgaggcatg gccgcgcgat ctgaaaggta tggcggctgt gcgtttcgac
481 cacgccctca tgtatggcga cgaattgccg gcgacctatg acctgttcac caaggtgctc
541 ggtttctatc tggccgaaca ggtgctggac gaaaatggca cgcgcgtcgc ccagtttctc
601 agtctgtcga ccaaggccca cgacgtggcc ttcattcacc atccggaaaa aggccgcctc
661 catcatgtgt ccttccacct cgaaacctgg gaagacttgc ttcgcgccgc cgacctgatc
721 tccatgaccg acacatctat cgatatcggc ccaacccgcc acggcctcac tcacggcaag
781 accatctact tcttcgaccc gtccggtaac cgcaacgaag tgttctgcgg gggagattac
841 aactacccgg accacaaacc ggtgacctgg accaccgacc agctgggcaa ggcgatcttt
901 taccacgacc gcattctcaa cgaacgattc atgaccgtgc tgacctgatg gtcccgggct
961 gcaggcatgc aagctgccca tcgagccgag aacgttatcg aagttagtca tgtgtaatcc
1021 cctcgtttga actttggatt aagcgtagat acacccttgg acaagccagt tggattcgga
1081 gacaagcaaa ttcagcctta aaaagggcga ggccctgcgg tggtggaaca ccgcagggcc
1141 tctaaccgct cgacgcgctg caccaaccag cccgcgaacg gctggcagcc agcgtaaggc
1201 gcggctcatc gggcggcgtt cgccacgatg tcctgcactt cgagccaagc ctcgaacacc
1261 tgctggtgtg cacgactcac ccggttgttg acaccgcgcg cggccgtgcg ggctcggtgg
1321 ggcggctctg tcgcccttgc cagcgtgagt agcgcgtacc tcacctcgcc caacaggtcg
1381 cacacagccg attcgtacgc cataaagcca ggtgagccca ccagctccgt aagttcgggc
1441 gctgtgtggc tcgtacccgc gcattcaggc ggcagggggt ctaacgggtc taaggcggcg
1501 tgtacggccg ccacagcggc tctcagcggc ccggaaacgt cctcgaaacg acgcatgtgt
1561 tcctcctggt tggtacaggt ggttgggggt gctcggctgt cgctggtgtt ccaccaccag
1621 ggctcgacgg gagagcgggg gagtgtgcag ttgtggggtg gcccctcagc gaaatatctg
1681 acttggagct cgtgtcggac catacaccgg tgattaatcg tggtctacta ccaagcgtga
1741 gccacgtcgc cgacgaattt gagcagctct ggctgccgta ctggccgctg gcaagcgacg
1801 atctgctcga ggggatctac cgccaaagcc gcgcgtcggc cctaggccgc cggtacatcg
1861 aggcgaaccc aacagcgctg gcaaacctgc tggtcgtgga cgtagaccat ccagacgcag
1921 cgctccgagc gctcagcgcc cgggggtccc atccgctgcc caacgcgatc gtgggcaatc
1981 gcgccaacgg ccacgcacac gcagtgtggg cactcaacgc ccctgttcca cgcaccgaat
2041 acgcgcggcg taagccgctc gcatacatgg cggcgtgcgc cgaaggcctt cggcgcgccg
2101 tcgacggcga ccgcagttac tcaggcctca tgaccaaaaa ccccggccac atcgcctggg
2161 aaacggaatg gctccactca gatctctaca cactcagcca catcgaggcc gagctcggcg
2221 cgaacatgcc accgccgcgc tggcgtcagc agaccacgta caaagcggct ccgacgccgc
2281 tagggcggaa ttgcgcactg ttcgattccg tcaggttgtg ggcctatcgt cccgccctca
2341 tgcggatcta cctgccgacc cggaacgtgg acggactcgg ccgcgcgatc tatgccgagt
2401 gccacacgcg aaacgccgaa ttctcgtgca acgacgtgtg tcccggaccg ctaccggaca
2461 gcgaggtccg cgccatcgcc aacagcattt ggcgttggat cacaaccaag tcgcgcattt
2521 gggcggacgg gatcgtggtc tacgaggcca cactcagtgc gcgccagtcg gccatctcgc
2581 ggaagggcgc agcagcgcgc acggcggcga gcacagttgc gcggcgcgca aagtccgcgt
2641 cagccatgga ggcattgcta tgagcgacgg ctacagcgac ggctacaacc ggcagccgac
2701 tgtccgcaaa aagcggcgcg tgaccgccgc cgaaggcgct cgaatcaccg gactatccga
2761 acgccacgtc gtccggctcg tggcgcagga acgcagcgag tggctcgccg agcaggctgc
2821 acgccgcgaa cgcatccgcg cctatcacga cgacgagggc cactcttggc cgcaaacggc
2881 caaacatttc gggctgcatc tggacaccgt taagcgactc ggctatcggg cgaggaaaga
2941 gcgtgcggca gaacaggaag cggctcaaaa ggcccacaac gaagccgaca atccaccgct
3001 gttctaacgc aattggggag cgggtgtcgc gggggttccg tggggggttc cgttgcaacg
3061 ggtcggacag gtaaaagtcc tggtagacgc tagttttctg gtttgggcca tgcctgtctc
3121 gttgcgtgtt tcgttgcgcc cgttttgaat accagccaga cgagacgggg ttctacgaat
3181 cttggtcgat accaagccat ttccgctgaa tatcggggag ctcaccgcca gaatcggtgg
3241 ttgtggtgat gtacgtggcg aactccgttg tagtgcctgt ggtggcatcc gtggccactc
3301 tcgttgcacg gttcgttgtg ccgttacagg ccccgttgac agctcaccga acgtagttaa
3361 aacatgctgg tcaaactagg tttaccaacg atacgagtca gctcatctag ggccagttct
3421 aggcgttgtt cgttgcgcgg ttcgttgcgc atgtttcgtg tggttgctag atggctccgc
3481 aaccacacgc ttcgaggttg agtgcttcca gcacgggcgc gatccagaag aacttcgtcg
3541 tgcgactgtc ctcgttaacg ctcgaagcga tgccgatcaa atcggctgtg ccgtcgcggt
3601 tcttgcggta cagcggcgcg cctgagtcgc catgtacggt gcgcagctga gcggtgatcg
3661 tgtacttggc agccacaacg accggcccac aggtctcgcc ggatttcatc ccgtacttgc
3721 agacctcggt gccgtacgga acatctttgc ggtccttcag caccgcacgc acggggtatc
3781 ttccgccgat tttgaacgaa gcgatcggtg ttcccggagc aaacgcaatc accccagcgt
3841 cggtgtagac cggcaataga tcatcgtcgc cggtaatcgt ggtgccttcg tctctcgacg
3901 caacgtaggg gggatccact agttctagag cggccgccac cgcggtggag ctccaattcg
3961 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
4021 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagggga
4081 attaattctg ctcgcgcagg ctgggtgcca agctctcggg taacatcaag gcccgatcct
4141 tggagccctt gccctcccgc acgatgatcg tgccgtgatc gaaatccaga tccttgaccc
4201 gcagttgcaa accctcactg atccgcatgc ccgttccata cagaagctgg gcgaacaaac
4261 gatgctcgcc ttccagaaaa ccgaggatgc gaaccacttc atccggggtc agcaccaccg
4321 gcaagcgccg cgacggccga ggtcttccga tctcctgaag ccagggcaga tccgtgcaca
4381 gcaccttgcc gtagaagaac agcaaggccg ccaatgcctg acgatgcgtg gagaccgaaa
4441 ccttgcgctc gttcgccagc caggacagaa atgcctcgac ttcgctgctg cccaaggttg
4501 ccgggtgacg cacaccgtgg aaacggatga aggcacgaac ccagttgaca taagcctgtt
4561 cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
4621 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
4681 ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
4741 tgatgttatg gagcagcaac gatgttacgc agcagcaacg atgttacgca gcagggcagt
4801 cgccctaaaa caaagttagg tggctcaagt atgggcatca ttcgcacatg taggctcggc
4861 cctgaccaag tcaaatccat gcgggctgct cttgatcttt tcggtcgtga gttcggagac
4921 gtagccacct actcccaaca tcagccggac tccgattacc tcgggaactt gctccgtagt
4981 aagacattca tcgcgcttgc tgccttcgac caagaagcgg ttgttggcgc tctcgcggct
5041 tacgttctgc ccaagtttga gcagccgcgt agtgagatct atatctatga tctcgcagtc
5101 tccggcgagc accggaggca gggcattgcc accgcgctca tcaatctcct caagcatgag
5161 gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag attacggtga cgatcccgca
5221 gtggctctct atacaaagtt gggcatacgg gaagaagtga tgcactttga tatcgaccca
5281 agtaccgcca cctaacaatt cgttcaagcc gagatcggct tcccggccgc ggagttgttc
5341 ggtaaattgt cacaacgccg cggccatcgg cattttcttt tgcgttttta tttgttaact
5401 gttaattgtc cttgttcaag gatgctgtct ttgacaacag atgttttctt gcctttgatg
5461 ttcagcagga agcttggcgc aaacgttgat tgtttgtctg cgtagaatcc tctgtttgtc
5521 atatagcttg taatcacgac attgtttcct ttcgcttgag gtacagcgaa gtgtgagtaa
5581 gtaaaggtta catcgttagg atcaagatcc atttttaaca caaggccagt tttgttcagc
5641 ggcttgtatg ggccagttaa agaattagaa acataaccaa gcatgtaaat atcgttagac
5701 gtaatgccgt caatcgtcat ttttgatccg cgggagtcag tgaacaggta ccatttgccg
5761 ttcattttaa agacgttcgc gcgttcaatt tcatctgtta ctgtgttaga tgcaatcagc
5821 ggtttcatca cttttttcag tgtgtaatca tcgtttagct caatcatacc gagagcgccg
5881 tttgctaact cagccgtgcg ttttttatcg ctttgcagaa gtttttgact ttcttgacgg
5941 aagaatgatg tgcttttgcc atagtatgct ttgttaaata aagattcttc gccttggtag
6001 ccatcttcag ttccagtgtt tgcttcaaat actaagtatt tgtggccttt atcttctacg
6061 tagtgaggat ctctcagcgt atggttgtcg cctgagctgt agttgccttc atcgatgaac
6121 tgctgtacat tttgatacgt ttttccgtca ccgtcaaaga ttgatttata atcctctaca
6181 ccgttgatgt tcaaagagct gtctgatgct gatacgttaa cttgtgcagt tgtcagtgtt
6241 tgtttgccgt aatgtttacc ggagaaatca gtgtagaata aacggatttt tccgtcagat
6301 gtaaatgtgg ctgaacctga ccattcttgt gtttggtctt ttaggataga atcatttgca
6361 tcgaatttgt cgctgtcttt aaagacgcgg ccagcgtttt tccagctgtc aatagaagtt
6421 tcgccgactt tttgatagaa catgtaaatc gatgtgtcat ccgcattttt aggatctccg
6481 gctaatgcaa agacgatgtg gtagccgtga tagtttgcga cagtgccgtc agcgttttgt
6541 aatggccagc tgtcccaaac gtccaggcct tttgcagaag agatattttt aattgtggac
6601 gaatcgaatt caggaacttg atatttttca tttttttgct gttcagggat ttgcagcata
6661 tcatggcgtg taatatggga aatgccgtat gtttccttat atggcttttg gttcgtttct
6721 ttcgcaaacg cttgagttgc gcctcctgcc agcagtgcgg tagtaaaggt taatactgtt
6781 gcttgttttg caaacttttt gatgttcatc gttcatgtct ccttttttat gtactgtgtt
6841 agcggtctgc ttcttccagc cctcctgttt gaagatggca agttagttac gcacaataaa
6901 aaaagaccta aaatatgtaa ggggtgacgc caaagtatac actttgccct ttacacattt
6961 taggtcttgc ctgctttatc agtaacaaac ccgcgcgatt tacttttcga cctcattcta
7021 ttagactctc gtttggattg caactggtct attttcctct tttgtttgat agaaaatcat
7081 aaaaggattt gcagactacg ggcctaaaga actaaaaaat ctatctgttt cttttcattc
7141 tctgtatttt ttatagtttc tgttgcatgg gcataaagtt gcctttttaa tcacaattca
7201 gaaaatatca taatatctca tttcactaaa taatagtgaa cggcaggtat atgtgatggg
7261 ttaaaaagga tcgatcctct agctagagtc gatcgacatc cgccctcacc gccaggaacg
7321 caaccgcagc ctcatcacgc cggcgcttct tggccgcgcg ggattcaacc cactcggcca
7381 gctcgtcggt gtagctcttt ggcatcgtct ctcgcctgtc ccctcagttc agtaatttcc
7441 tgcatttgcc tgtttccagt cggtagatat tccacaaaac agcagggaag cagcgctttt
7501 ccgctgcata accctgcttc ggggtcatta tagcgatttt ttcggtatat ccatcctttt
7561 tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac
7621 ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc gagcgggtgt tccttcttca
7681 ctgtccctta ttcgcacctg gcggtgctca acgggaatcc tgctctgcga ggctggccgg
7741 ctaccgccgg cgtaacagat gagggcaagc ggatggctga tgaaaccaag ccaaccagga
7801 agggcagccc acctatcaag gtgtactgcc ttccagacga acgaagagcg attgaggaaa
7861 aggcggcggc ggccggcatg agcctgtcgg cctacctgct ggccgtcggc cagggctaca
7921 aaatcacggg cgtcgtggac tatgagcacg tccgcgagct ggcccgcatc aatggcgacc
7981 tgggccgcct gggcggcctg ctgaaactct ggctcaccga cgacccgcgc acggcgcggt
8041 tcggtgatgc cacgatcctc gccctgctgg cgaagatcga ctctagctag aaatatttta
8101 tctgattaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg
8161 aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt
8221 tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc
8281 cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc
8341 agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg
8401 cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac
8461 ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc
8521 ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg gaaacgcctg
8581 gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg
8641 cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc gcggccctct cacttccctg
8701 ttaagtatct tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc catttccgct
8761 cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga atatatcctg
8821 tatcacatat tctgctgacg caccggtgca gccttttttc tcctgccaca tgaagcactt
8881 cactgacacc ctcatcagtg ccaacatagt aagccagtat acactccgct agcgctgatg
8941 tccggcggtg cttttgccgt tacgcaccac cccgtcagta gctgaacagg agggacagaa
9001 ttaattcccc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
9061 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc tcccggctcg
9121 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
9181 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctgggta ccgggccccc
9241 cctcgaggtc gacggtatcg ataagcttga tatcgaattc cctagacggt gaccacaacg
9301 cgcccgcttt gatcggggac gtctgcggcc gaccatttac gggtcttgtt gtcgttggcg
9361 gtcatgggcc gaacatactc acccggatcg gagggccgag gacaaggtcg aacgaggggc
9421 atgacccggt gcggggcttc ttgcactcgg cataggcgag tgctaagaat aacgttggca
9481 ctcgcgaccg gtgagtgcta ggtcgggacg gtgaggccag gcccgtcgtc gcagcgagtg
9541 gcagcgagga caacttgagc cgtccgtcgc gggcactgcg cccggccagc gtaagtagcg
9601 gggttgccgt cacccggtga cccccgtttc atccccgatc cggaggaatc acttcgcaat
9661 ggccaagaca attgcggatc c
//
