LOCUS Exported 9111 bp ds-DNA circular SYN 13-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9111)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Tuesday, September 13, 2016 from SnapGene Viewer 3.2.1
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..9111
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1219..1848
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/note="pVS1 StaA"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 2277..3350
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/note="pVS1 RepA"
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 3416..3610
/note="pVS1 oriV"
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 3954..4094
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(4280..4868)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5139..5798)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
misc_feature 6326..6350
/note="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 6428..6602
/note="CaMV poly(A) signal"
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(6642..7667)
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/note="HygR"
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
promoter complement(7735..8412)
/note="CaMV 35S promoter (enhanced)"
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 8603..8624
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8639..8669
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 8677..8693
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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 8701..8717
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 8713..8946
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
misc_feature 8727..8783
/note="MCS"
/note="pUC18/19 multiple cloning site"
primer_bind complement(8787..8803)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 9006..9030
/note="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
ORIGIN
1 catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc ctccgctgct
61 atagtgcagt cggcttctga cgttcagtgc agccgtcttc tgaaaacgac atgtcgcaca
121 agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt
181 gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac gccatgaaca
241 agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac caggacttga
301 ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc gagaagatca
361 ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta cgccctggcg
421 acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac ctactggaca
481 ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg
541 acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg
601 agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg
661 tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc cgcgagctga
721 tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg catcgctcga
781 ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg
841 gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac
901 gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt ttttcattac
961 cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacgt
1021 ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg
1081 gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt
1141 tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca
1201 aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc
1261 aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg
1321 ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa
1381 ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc
1441 cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg
1501 atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc
1561 accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa
1621 gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag
1681 gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac
1741 ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc
1801 cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta
1861 aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca
1921 gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc
1981 agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca
2041 ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa
2101 atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc
2161 accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc
2221 tgggttgtct gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga
2281 gcggtcgcaa accatccggc ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg
2341 agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg
2401 gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag
2461 ccggtgcgcc gtcgattagg aagccgccca agggcgacga gcaaccagat tttttcgttc
2521 cgatgctcta tgacgtgggc acccgcgata gtcgcagcat catggacgtg gccgttttcc
2581 gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg ctacgagctt ccagacgggc
2641 acgtagaggt ttccgcaggg ccggccggca tggccagtgt gtgggattac gacctggtac
2701 tgatggcggt ttcccatcta accgaatcca tgaaccgata ccgggaaggg aagggagaca
2761 agcccggccg cgtgttccgt ccacacgttg cggacgtact caagttctgc cggcgagccg
2821 atggcggaaa gcagaaagac gacctggtag aaacctgcat tcggttaaac accacgcacg
2881 ttgccatgca gcgtacgaag aaggccaaga acggccgcct ggtgacggta tccgagggtg
2941 aagccttgat tagccgctac aagatcgtaa agagcgaaac cgggcggccg gagtacatcg
3001 agatcgagct agctgattgg atgtaccgcg agatcacaga aggcaagaac ccggacgtgc
3061 tgacggttca ccccgattac tttttgatcg atcccggcat cggccgtttt ctctaccgcc
3121 tggcacgccg cgccgcaggc aaggcagaag ccagatggtt gttcaagacg atctacgaac
3181 gcagtggcag cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt
3241 caaatgacct gccggagtac gatttgaagg aggaggcggg gcaggctggc ccgatcctag
3301 tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc cggttcctaa tgtacggagc
3361 agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg aaaaggtctc tttcctgtgg
3421 atagcacgta cattgggaac ccaaagccgt acattgggaa ccggaacccg tacattggga
3481 acccaaagcc gtacattggg aaccggtcac acatgtaagt gactgatata aaagagaaaa
3541 aaggcgattt ttccgcctaa aactctttaa aacttattaa aactcttaaa acccgcctgg
3601 cctgtgcata actgtctggc cagcgcacag ccgaagagct gcaaaaagcg cctacccttc
3661 ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc tatcgcggcc gctggccgct
3721 caaaaatggc tggcctacgg ccaggcaatc taccagggcg cggacaagcc gcgccgtcgc
3781 cactcgaccg ccggcgccca catcaaggca ccctgcctcg cgcgtttcgg tgatgacggt
3841 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
3901 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
3961 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
4021 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
4081 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
4141 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
4201 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
4261 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
4321 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
4381 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
4441 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
4501 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
4561 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
4621 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
4681 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
4741 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
4801 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
4861 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
4921 cacgttaagg gattttggtc atgcatgata tatctcccaa tttgtgtagg gcttattatg
4981 cacgcttaaa aataataaaa gcagacttga cctgatagtt tggctgtgag caattatgtg
5041 cttagtgcat ctaatcgctt gagttaacgc cggcgaagcg gcgtcggctt gaacgaattt
5101 ctagctagag gatcgcacca ataactgcct taaaaaaatt acgccccgcc ctgccactca
5161 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
5221 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
5281 attgtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
5341 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
5401 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
5461 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
5521 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg
5581 aactccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
5641 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
5701 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
5761 atatcaacgg tggtatatcc agtgattttt ttctccatga tgtttaactt tgttttaggg
5821 cgactgccct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg
5881 taacgcgctt gctgcttgga tgcccgaggc atagactgta ccccaaaaaa acatgtcata
5941 acaagaagcc atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct
6001 ggaccagttg cgtgacggca gttacgctac ttgcattaca gcttacgaac cgaacgaggc
6061 ttatgtccac tgggttcgtg cccgaattga tcacaggcag caacgctctg tcatcgttac
6121 aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg
6181 ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc
6241 tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc
6301 ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac aaattgacgc
6361 ttagacaact taataacaca ttgcggacgt ttttaatgta ctgaattaac gccgaattaa
6421 ttcgggggat ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat
6481 agaagtattt tacaaataca aatacatact aagggtttct tatatgctca acacatgagc
6541 gaaaccctat aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga
6601 aactcgagct tgtcgatcga cagatccggt cggcatctac tctatttctt tgccctcgga
6661 cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag
6721 acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg
6781 attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc
6841 tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagtcgtgg cgatcctgca
6901 agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc
6961 ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag
7021 tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg
7081 tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc
7141 tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga
7201 tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg
7261 aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catagcctcc
7321 gcgaccggtt gtagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc
7381 tgtgcacggc gggagatgca ataggtcagg ctctcgctaa actccccaat gtcaagcact
7441 tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa
7501 ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa
7561 gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg
7621 atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatatc tcattgcccc
7681 ccgggatctg cgaaagctcg agagagatag atttgtagag agagactggt gatttcagcg
7741 tgtcctctcc aaatgaaatg aacttcctta tatagaggaa ggtcttgcga aggatagtgg
7801 gattgtgcgt catcccttac gtcagtggag atatcacatc aatccacttg ctttgaagac
7861 gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga
7921 ccactgtcgg cagaggcatc ttgaacgata gcctttcctt tatcgcaatg atggcatttg
7981 taggtgccac cttccttttc tactgtcctt ttgatgaagt gacagatagc tgggcaatgg
8041 aatccgagga ggtttcccga tattaccctt tgttgaaaag tctcaatagc cctttggtct
8101 tctgagactg tatctttgat attcttggag tagacgagag tgtcgtgctc caccatgtta
8161 tcacatcaat ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc
8221 ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag aggcatcttg aacgatagcc
8281 tttcctttat cgcaatgatg gcatttgtag gtgccacctt ccttttctac tgtccttttg
8341 atgaagtgac agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt
8401 tgaaaagtct caatagccct ttggtcttct gagactgtat ctttgatatt cttggagtag
8461 acgagagtgt cgtgctccac catgttggca agctgctcta gccaatacgc aaaccgcctc
8521 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
8581 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
8641 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
8701 caggaaacag ctatgaccat gattacgaat tcgagctcgg tacccgggga tcctctagag
8761 tcgacctgca ggcatgcaag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa
8821 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
8881 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
8941 tgctagagca gcttgagctt ggatcagatt gtcgtttccc gccttcagtt taaactatca
9001 gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataac
9061 ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt g
//