LOCUS       Exported                7146 bp ds-DNA     circular SYN 08-JUN-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7146)
  AUTHORS   MRC LMB
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 19, 2019 from SnapGene Viewer 4.3.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7146
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     907..930
                     /label=JBPR64
     CDS             913..1473
                     /codon_start=1
                     /label=Cdc42
                     /translation="MQTIKCVVVGDGAVGKNCLLISYTTNKFPSEYVPTVFDNYAVTVM
                     IGGEPYTLGLFDTAGQEDYDRLRPLSYPQTDVFLVCFSVVSPSSFENVKEKWVPEITHH
                     CPKTPFLLVGTQIDLRDDPSTIEKLAKNKQKPITPETAEKLARDLKAVKYVECSALTQR
                     GLKNVFDEAILAALEPPETQPKRK"
     primer_bind     940..984
                     /label=JBPR66
     primer_bind     complement(940..984)
                     /label=JBPR67
     primer_bind     complement(1456..1481)
                     /label=JBPR65
     CDS             1498..2463
                     /codon_start=1
                     /label=BirA*
                     /translation="MDKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLR
                     DWGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKS
                     GDACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLR
                     KLGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQ
                     GWITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLII
                     GDKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     primer_bind     1498..1519
                     /label=JBPR73 (seq primer)
     primer_bind     1498..1518
                     /label=JBPR1
     primer_bind     2422..2443
                     /label=JBPR3
     primer_bind     complement(2440..2463)
                     /label=JBPR6
     primer_bind     complement(2446..2463)
                     /label=JBPR2
     CDS             2494..2520
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2746..2970
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3016..3444
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3458..3787
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3638..3773
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3854..4648
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4822..4943
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4992..5008)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5016..5032
                     /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)."
     promoter        complement(5040..5070)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5085..5106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5394..5979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6150..7010)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7011..7115)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 aagcttaccg gtatgcagac aattaagtgt gttgttgtgg gcgatggtgc tgttggtaaa
      961 aactgtctcc tgatatccta cacaacaaac aaatttccat cggaatatgt accgactgtt
     1021 tttgacaact atgcagtcac agttatgatt ggtggagaac catatactct tggacttttt
     1081 gatactgcag ggcaagagga ttatgacaga ttacgaccgc tgagttatcc acaaacagat
     1141 gtatttctag tctgtttttc agtggtctct ccatcttcat ttgaaaacgt gaaagaaaag
     1201 tgggtgcctg agataactca ccactgtcca aagactcctt tcttgcttgt tgggactcaa
     1261 attgatctca gagatgaccc ctctactatt gagaaacttg ccaagaacaa acagaagcct
     1321 atcactccag agactgctga aaagctggcc cgtgacctga aggctgtcaa gtatgtggag
     1381 tgttctgcac ttacacagag aggtctgaag aatgtgtttg atgaggctat cctagctgcc
     1441 ctcgagcctc cggaaactca acccaaaagg aaggaattcg gagccggagc tggtaccatg
     1501 gacaaggaca acaccgtgcc cctgaagctg atcgccctgc tggccaacgg cgagttccac
     1561 tctggcgagc agctgggaga gaccctggga atgagcagag ccgccatcaa caagcacatc
     1621 cagacactga gagactgggg agtggacgtg ttcaccgtgc ctggcaaggg ctacagcctg
     1681 cctgagccta tccagctgct gaacgccaag cagatcctgg gacagctgga tggcggaagc
     1741 gtggccgtgc tgcctgtgat cgactccacc aatcagtacc tgctggacag aatcggagag
     1801 ctgaagtccg gcgacgcctg catcgccgag taccagcagg ctggcagagg aggcagagga
     1861 cggaagtggt tcagcccatt cggagccaac ctgtacctgt ccatgttctg gagactggag
     1921 cagggacctg ctgctgccat cggactgagt ctggtgatcg gaatcgtgat ggccgaggtg
     1981 ctgagaaagc tgggagccga caaggtgaga gtgaagtggc ctaatgacct gtacctccag
     2041 gaccgcaagc tggctggcat cctggtggag ctgacaggca agacaggcga tgccgctcag
     2101 atcgtgatcg gagccggaat caacatggcc atgagaagag tggaggagag cgtggtgaac
     2161 cagggctgga tcaccctgca ggaggctggc atcaacctgg accggaacac cctggccgcc
     2221 atgctgatca gagagctgag agccgctctg gagctgttcg agcaggaggg actggctcct
     2281 tacctgagca gatgggagaa gctggacaac ttcatcaaca gacctgtgaa gctgatcatc
     2341 ggcgacaagg aaatcttcgg catctccaga ggaatcgaca agcagggagc tctgctgctg
     2401 gagcaggacg gaatcatcaa gccctggatg ggcggagaaa tctccctgag aagcgcagag
     2461 aagggatccg tcgacggagc cggtgctggc gcatacccct acgacgtgcc cgactacgca
     2521 ggctccgcag ttccagcggt agaaatcacc cacaccttct gggaaaggaa cctgccctct
     2581 gtttctggcc tgctgaagat cattggattt tccacatcag taactgccct ggggtttgtg
     2641 ctgtacaaat acaagctcct gccacggtct tgacccgggg atatccagca cagtggcggc
     2701 cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     2761 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2821 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2881 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2941 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3001 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3061 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3121 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3181 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3241 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3301 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3361 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3421 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3481 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3541 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3601 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3661 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3721 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3781 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     3841 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3901 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3961 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4021 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4081 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4141 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4201 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4261 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4321 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4381 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4441 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4501 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4561 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4621 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4681 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4741 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4801 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4861 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4921 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4981 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5041 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5101 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5161 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5221 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5281 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5341 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5401 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5461 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5521 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5581 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5641 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5701 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5761 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5821 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5881 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt
     5941 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6001 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6061 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6121 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6181 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6241 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6301 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6361 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6421 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6481 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6541 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6601 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6661 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6721 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6781 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6841 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6901 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6961 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7021 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7081 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7141 gacgtc
//
