LOCUS Exported 5704 bp ds-DNA circular SYN 01-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..5704
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 183..211
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 219..235
/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)."
CDS 258..911
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
CDS 918..935
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 969..1682
/codon_start=1
/product="monomeric derivative of red fluorescent protein
from Entacmaea quadricolor (Merzlyak et al., 2007)"
/label=TagRFP
/note="mammalian codon-optimized"
/translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
ARYCDLPSKLGHKLN"
promoter 2007..2111
/gene="bla"
/label=AmpR promoter
CDS 2112..2972
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3143..3731
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 3975..4052
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 4053..5135
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
CDS 5258..5704
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGI"
ORIGIN
1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
901 atcctccaaa atcggatctg gttccgcgtg gatccccggg aatttccggt ggtggtggtg
961 gaattctaat ggtgtctaag ggcgaagagc tgattaagga gaacatgcac atgaagctgt
1021 acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc
1081 cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg
1141 ccttcgacat cctggctacc agcttcatgt acggcagcag aaccttcatc aaccacaccc
1201 agggcatccc cgacttcttt aagcagtcct tccctgaggg cttcacatgg gagagagtca
1261 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
1321 gcctcatcta caacgtcaag atcagagggg tgaacttccc atccaacggc cctgtgatgc
1381 agaagaaaac actcggctgg gaggccaaca ccgagatgct gtaccccgct gacggcggcc
1441 tggaaggcag aagcgacatg gccctgaagc tcgtgggcgg gggccacctg atctgcaact
1501 tcaagaccac atacagatcc aagaaacccg ctaagaacct caagatgccc ggcgtctact
1561 atgtggacca cagactggaa agaatcaagg aggccgacaa agagacctac gtcgagcagc
1621 acgaggtggc tgtggccaga tactgcgacc tccctagcaa actggggcac aaacttaatt
1681 gactcgagct caagcttatt catcgtgact gactgacgat ctgcctcgcg cgtttcggtg
1741 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
1801 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
1861 gcgcagccat gacccagtca cgtagcgata gcggagtgta taattcttga agacgaaagg
1921 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
1981 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
2041 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
2101 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
2161 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
2221 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
2281 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
2341 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
2401 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
2461 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
2521 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
2581 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
2641 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
2701 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
2761 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
2821 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
2881 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
2941 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
3001 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
3061 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
3121 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
3181 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
3241 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
3301 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
3361 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
3421 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
3481 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
3541 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
3601 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
3661 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
3721 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
3781 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
3841 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
3901 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
3961 accgcataaa ttccgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc
4021 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
4081 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
4141 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
4201 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
4261 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
4321 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
4381 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
4441 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
4501 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
4561 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
4621 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
4681 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
4741 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
4801 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg
4861 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
4921 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
4981 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
5041 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
5101 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
5161 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
5221 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cggattcact
5281 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
5341 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
5401 ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc cggcaccaga
5461 agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg tcgtcgtccc
5521 ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtaa cctatcccat
5581 tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact cgctcacatt
5641 taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg atggcgttgg
5701 aatt
//