LOCUS       Exported                7356 bp ds-DNA     circular SYN 27-JUL-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7356)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Jul 27, 2021 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7356
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             85..540
                     /codon_start=1
                     /product="lysozyme from bacteriophage T7"
                     /label=T7 lysozyme
                     /note="inhibitor of T7 RNA polymerase"
                     /translation="MARVQFKQRESTDAIFVHCSATKPSQNVGVREIRQWHKEQGWLDV
                     GYHFIIKRDGTVEAGRDEMAVGSHAKGYNHNSIGVCLVGGIDDKGKFDANFTPAQMQSL
                     RSLLVTLLAKYEGAVLRAHHEVAPYACPSFDLKRWWEKNELVTSDRG"
     primer_bind     375..392
                     /label=pCysDisCo Seq. 8
                     /note="oABB008"
     primer_bind     complement(587..615)
                     /label=CyDisCo_loyid_rrnC_rev
     terminator      610..657
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(630..669)
                     /label=pLemo_rev
                     /note="oABB012"
     primer_bind     661..692
                     /label=CyDisCo_J231xx_fwd_insert
                     /note="oABB015"
     primer_bind     complement(673..748)
                     /label=pTac-in-pLemo-CyDisCo_rev
     promoter        693..721
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     promoter        693..694
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    729..745
                     /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     739..821
                     /label=pTac-in-pLemo-CyDisCo_fwd
     primer_bind     complement(767..802)
                     /label=Rv-Erv1p-RBS-change
     primer_bind     802..866
                     /label=J231xx in cydisco_fwd
     RBS             805..827
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     823..859
                     /label=Fw-Erv1p-RBS-change
     misc_feature    834..1406
                     /label=Erv1p
     primer_bind     835..868
                     /label=CyDisCo_wo ptac
     primer_bind     946..965
                     /label=pCysDisCo Seq. 2
                     /note="oABB002"
     RBS             1431..1453
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1459..2940
                     /codon_start=1
                     /label=hPDI
                     /translation="MDAPEEEDHVLVLRKSNFAEALAAHKYLLVEFYAPWCGHCKALAP
                     EYAKAAGKLKAEGSEIRLAKVDATEESDLAQQYGVRGYPTIKFFRNGDTASPKEYTAGR
                     EADDIVNWLKKRTGPAATTLPDGAAAESLVESSEVAVIGFFKDVESDSAKQFLQAAEAI
                     DDIPFGITSNSDVFSKYQLDKDGVVLFKKFDEGRNNFEGEVTKENLLDFIKHNQLPLVI
                     EFTEQTAPKIFGGEIKTHILLFLPKSVSDYDGKLSNFKTAAESFKGKILFIFIDSDHTD
                     NQRILEFFGLKKEECPAVRLITLEEEMTKYKPESEELTAERITEFCHRFLEGKIKPHLM
                     SQELPEDWDKQPVKVLVGKNFEDVAFDEKKNVFVEFYAPWCGHCKQLAPIWDKLGETYK
                     DHENIVIAKMDSTANEVEAVKVHSFPTLKFFPASADRTVIDYNGERTLDGFKKFLESGG
                     QDGAGDDDDLEDLEEAEEPDMEEDDDQKAVKDEL*"
     primer_bind     1584..1601
                     /label=pCysDisCo Seq. 3
                     /note="oABB003"
     primer_bind     2217..2235
                     /label=pCysDisCo Seq. 4
                     /note="oABB004"
     primer_bind     2886..2902
                     /label=pCysDisCo Seq. 5
                     /note="oABB005"
     CDS             2993..3010
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3054..3130)
                     /label=CyDisCo_loyid_rrnC_rev
     terminator      3077..3082
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3122..3157
                     /label=pLEMO-BB_fwd
                     /note="oABB020"
     primer_bind     complement(3166..3188)
                     /label=pCysDisCo Seq. 1
                     /note="oABB001"
     CDS             complement(3232..3891)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3892..3994)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(4520..5064)
                     /direction=LEFT
                     /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     complement(5128..5167)
                     /label=pCyCisCo_rev
                     /note="oABB010"
     CDS             complement(5395..6243)
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     primer_bind     5395..5418
                     /label=pLemo_fwd
                     /note="oABB011"
     CDS             complement(6317..7153)
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter"
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     promoter        join(7299..7356,1..61)
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
ORIGIN
        1 gcccattttc ctgtcagtaa cgagaaggtc gcgaattcag gcgcttttta gactggtcgt
       61 aatgaaattc aggaggtggt cgacatggct cgtgtacagt ttaaacaacg tgaatctact
      121 gacgcaatct ttgttcactg ctcggctacc aagccaagtc agaatgttgg tgtccgtgag
      181 attcgccagt ggcacaaaga gcagggttgg ctcgatgtgg gataccactt tatcatcaag
      241 cgagacggta ctgtggaggc aggacgagat gagatggctg taggctctca cgctaagggt
      301 tacaaccaca actctatcgg cgtctgcctt gttggtggta tcgacgataa aggtaagttc
      361 gacgctaact ttacgccagc ccaaatgcaa tcccttcgct cactgcttgt cacactgctg
      421 gctaagtacg aaggcgctgt gcttcgcgcc catcatgagg tggcgccgta cgcttgccct
      481 tcgttcgacc ttaagcgttg gtgggagaag aacgaactgg tcacttctga ccgtggataa
      541 ggatccccgc gccctcatcc gaaagggcgt attcatatgc cctaggctgc tgccaccgct
      601 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgcat
      661 aacggttctg gcaaatattc tgaaatgagc tgttgacaat taatcatcgg ctcgtataat
      721 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agccagtccg tttaggtgtt
      781 ttcacgagct ctagaaataa ttatcagcag gacgcactga ccaggaggta catatgaaag
      841 ccatcgataa aatgaccgac aatccgcctc aagagggtct gagtggtcgt aaaatcatct
      901 atgatgagga cggcaaacca tgtcgcagct gcaataccct gctggacttc cagtatgtta
      961 ccgggaaaat ctcaaatggc ctgaaaaacc tgagcagcaa tggtaaactg gcgggtacag
     1021 gtgctctgac tggggaagca tctgaactga tgccgggttc tcgtacctat cgtaaagtcg
     1081 atcctccgga tgttgaacag ctgggacgtt cttcatggac actgctgcat agcgtagcag
     1141 cctcttatcc agctcaacca accgatcagc aaaaaggcga aatgaaacag ttcctgaaca
     1201 tcttcagcca catctatccg tgtaactggt gcgctaaaga cttcgaaaaa tatatccgtg
     1261 agaatgcccc tcaagttgaa tcacgtgagg agctgggtcg ctggatgtgt gaggcccata
     1321 acaaagtgaa caaaaagctg cgcaaaccga agttcgactg taacttctgg gaaaaacgct
     1381 ggaaagatgg ttgggatgag taataaggat ccgaattctc tagtaataat tttgtttaac
     1441 tttaagaagg agatacatat ggatgcccca gaggaggaag atcacgtcct ggttctgcgt
     1501 aaaagcaact tcgctgaagc actggcagct cacaaatatc tgctggtcga gttctatgct
     1561 ccgtggtgtg gtcattgcaa agccctggct ccggaatatg ctaaagcagc cggcaaactg
     1621 aaagctgagg gcagtgaaat tcgtctggcc aaagtggacg ctaccgaaga atcagatctg
     1681 gcacaacagt atggtgttcg tggttatccg actatcaaat ttttccgtaa cggcgataca
     1741 gcaagcccta aagagtatac cgctggccgt gaagctgatg atatcgtgaa ctggctgaaa
     1801 aaacgtacag gtccggcggc aacgactctg cctgatggtg ccgctgccga gtcactggta
     1861 gaatcatccg aagtggccgt gattggcttc tttaaagacg tggagagcga ttcagcaaaa
     1921 cagttcctgc aagcagctga agcgattgat gacatcccgt ttggtattac gagcaatagc
     1981 gacgtgttct ccaaatatca actggacaaa gacggtgtgg ttctgttcaa aaaattcgac
     2041 gaaggccgta acaactttga aggtgaggtg accaaagaaa acctgctgga ctttatcaaa
     2101 cacaatcaac tgccgctggt gattgagttc accgaacaga cagctccgaa aatctttggc
     2161 ggcgagatca aaacccacat tctgctgttt ctgcctaaaa gtgtgtccga ctatgacggc
     2221 aaactgagca acttcaaaac cgccgctgag agctttaaag gcaaaatcct gttcatcttc
     2281 atcgacagcg atcataccga caaccagcgt attctggagt tttttggcct gaaaaaagag
     2341 gaatgtccgg ccgttcgcct gattacactg gaagaggaga tgacgaaata taaaccagaa
     2401 agcgaggagc tgacagctga acgtattacc gagttctgcc accgttttct ggaaggcaaa
     2461 atcaaacctc atctgatgtc ccaagaactg ccagaagatt gggataaaca gcctgtgaaa
     2521 gtgctggtag gcaaaaactt cgaggatgtg gcgttcgacg agaaaaaaaa cgtgtttgtg
     2581 gagttttatg ccccttggtg tggacactgc aaacagctgg caccgatttg ggataaactg
     2641 ggcgaaacct ataaagatca tgaaaacatt gttattgcca aaatggacag caccgccaat
     2701 gaagtcgaag ccgtgaaagt tcattcgttt ccgaccctga aattctttcc tgccagcgcc
     2761 gatcgtactg tgatcgatta taacggggag cgtaccctgg atggttttaa aaaattcctg
     2821 gagagcggtg gtcaagatgg tgccggtgat gatgacgatc tggaggatct ggaagaggct
     2881 gaagaaccgg atatggagga ggatgacgat cagaaagcag tcaaagacga gctgtgataa
     2941 ggatccgaat tcactagtga gctccgtcga caagcttgcg gccgcactcg agcaccacca
     3001 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     3061 caccgctgag caataactag cagataaaaa aaatccttag ctttcgctaa ggatgatttc
     3121 tactggttat ggtcgaattt gctttcgaat ttctgccatt catccgctta ttatcactta
     3181 ttcaggcgta gcaccaggcg tttaagggca ccaataactg ccttaaaaaa attacgcccc
     3241 gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc
     3301 atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt
     3361 ataatatttg cccatagtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa
     3421 atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa
     3481 ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg
     3541 tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg
     3601 ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt
     3661 cattgccata cggaactccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc
     3721 cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg
     3781 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
     3841 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc
     3901 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     3961 atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc aaaagttggc
     4021 ccagggcttc ccggtatcaa cagggacacc aggatttatt tattctgcga agtgatcttc
     4081 cgtcacaggt atttattcgg cgcaaagtgc gtcgggtgat gctgccaact tactgattta
     4141 gtgtatgatg gtgtttttga ggtgctccag tggcttctgt ttctatcagc tgtccctcct
     4201 gttcagctac tgacggggtg gtgcgtaacg gcaaaagcac cgccggacat cagcgctagc
     4261 ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag tgcttcatgt
     4321 ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag gatatattcc
     4381 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct
     4441 tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga
     4501 gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct
     4561 gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4621 tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct
     4681 gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct
     4741 ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta
     4801 actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg
     4861 gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg
     4921 acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct
     4981 cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt
     5041 acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa atatttctag
     5101 gtttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag
     5161 ttgtaattct catgttagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     5221 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     5281 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5341 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg cccgataagc ttaattaatc
     5401 tttctgcgaa ttgagatgac gccactggct gggcgtcatc ccggtttccc gggtaaacac
     5461 caccgaaaaa tagttactat cttcaaagcc acattcggtc gaaatatcac tgattaacag
     5521 gcggctatgc tggagaagat attgcgcatg acacactctg acctgtcgca gatattgatt
     5581 gatggtcatt ccagtctgct ggcgaaattg ctgacgcaaa acgcgctcac tgcacgatgc
     5641 ctcatcacaa aatttatcca gcgcaaaggg acttttcagg ctagccgcca gccgggtaat
     5701 cagcttatcc agcaacgttt cgctggatgt tggcggcaac gaatcactgg tgtaacgatg
     5761 gcgattcagc aacatcacca actgcccgaa cagcaactca gccatttcgt tagcaaacgg
     5821 cacatgctga ctactttcat gctcaagctg accgataacc tgccgcgcct gcgccatccc
     5881 catgctacct aagcgccagt gtggttgccc tgcgctggcg ttaaatcccg gaatcgcccc
     5941 ctgccagtca agattcagct tcagacgctc cgggcaataa ataatattct gcaaaaccag
     6001 atcgttaacg gaagcgtagg agtgtttatc gtcagcatga atgtaaaaga gatcgccacg
     6061 ggtaatgcga taagggcgat cgttgagtac atgcaggcca ttaccgcgcc agacaatcac
     6121 cagctcacaa aaatcatgtg tatgttcagc aaagacatct tgcggataac ggtcagccac
     6181 agcgactgcc tgctggtcgc tggcaaaaaa atcatctttg agaagtttta actgatgcgc
     6241 caccgtggct acctcggcca gagaacgaag ttgattattc gcaatatggc gtacaaatac
     6301 gttgagaaga ttcgcgttat tgcagaaagc catcccgtcc ctggcgaata tcacgcggtg
     6361 accagttaaa ctctcggcga aaaagcgtcg aaaagtggtt actgtcgctg aatccacagc
     6421 gataggcgat gtcagtaacg ctggcctcgc tgtggcgtag cagatgtcgg gctttcatca
     6481 gtcgcaggcg gttcaggtat cgctgaggcg tcagtcccgt ttgctgctta agctgccgat
     6541 gtagcgtacg cagtgaaaga gaaaattgat ccgccacggc atcccaattc acctcatcgg
     6601 caaaatggtc ctccagccag gccagaagca agttgagacg tgatgcgctg ttttccaggt
     6661 tctcctgcaa actgctttta cgcagcaaga gcagtaattg cataaacaag atctcgcgac
     6721 tggcggtcga gggtaaatca ttttcccctt cctgctgttc catctgtgca accagctgtc
     6781 gcacctgctg caatacgctg tggttaacgc gccagtgaga cggatactgc ccatccagct
     6841 cttgtggcag caactgattc agcccggcga gaaactgaaa tcgatccggc gagcgataca
     6901 gcacattggt cagacacaga ttatcggtat gttcatacag atgccgatca tgatcgcgta
     6961 cgaaacagac cgtgccaccg gtgatggtat agggctgccc attaaacaca tgaatacccg
     7021 tgccatgttc gacaatcaca atttcatgaa aatcatgatg atgttcagga aaatccgcct
     7081 gcgggagccg gggttctatc gccacggacg cgttaccaga cggaaaaaaa tccacactat
     7141 gtaatacggt catactggcc tcctgatgtc gtcaacacgg cgaaatagta atcacgaggt
     7201 caggttctta ccttaaattt tcgacggaaa accacgtaaa aaacgtcgat ttttcaagat
     7261 acagcgtgaa ttttcaggaa atgcggtgag catcacatca ccacaattca gcaaattgtg
     7321 aacatcatca cgttcatctt tccctggttg ccaatg
//
