LOCUS       Exported                8769 bp ds-DNA     circular SYN 09-DEC-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8769)
  AUTHORS   sa
  TITLE     Direct Submission
  JOURNAL   Exported Feb 6, 2018 from SnapGene 4.1.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8769
                     /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"
     misc_feature    907..4617
                     /label=h13LB
     CDS             4597..4617
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4633..4674
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4684..4701
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4730..4954
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5000..5428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5442..5772
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5623..5758
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5820..5867
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5886..6284
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6442..6563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6612..6628)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6636..6652
                     /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(6660..6690)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6705..6726
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7014..7602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7773..8633)
                     /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(8634..8738)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gccaccatgt caaagctgga gaaattcacc aactgttata gcctgtctaa gaccctgcgc
      961 ttcaaggcaa tcccagtggg caagacacaa gagaacattg acaacaaacg gctcctggtg
     1021 gaggatgaga agagggctga agattacaag ggcgttaaga agctgctgga taggtactat
     1081 ctgtcattca tcaacgatgt cctccacagt atcaagctga agaatctgaa caattacatt
     1141 tctctgttcc ggaagaagac acggaccgag aaggagaaca aagagctgga gaatctggag
     1201 atcaacctga ggaaagaaat agctaaggct ttcaaaggga acgagggtta caagtccctg
     1261 ttcaagaaag acattatcga gactattctg cctgagttcc tggacgataa agatgagatc
     1321 gccctcgtca attccttcaa tgggtttacc acagccttta ccggcttctt cgacaataga
     1381 gagaatatgt tctctgaaga ggccaaatcc actagcatcg cctttcgctg cataaacgag
     1441 aacctgacta ggtacatcag caatatggac atctttgaga aagtcgatgc catattcgac
     1501 aaacatgagg tgcaggagat taaggagaag atcctgaact cagattacga tgtcgaagat
     1561 ttcttcgagg gagagttctt caacttcgtg ctcacacaag agggcattga tgtgtacaat
     1621 gcaatcattg gagggttcgt gacagagagt ggcgagaaga taaagggcct gaacgagtat
     1681 atcaacctct acaaccagaa aaccaagcag aaactgccta agttcaagcc actgtacaaa
     1741 caagtgctct cagataggga aagcctgagc ttctacggtg aagggtatac atcagatgaa
     1801 gaagtgctcg aagtgttccg caacaccctc aataagaaca gtgaaatctt ctcttcaatc
     1861 aagaagctgg agaaactgtt caagaatttc gatgagtact cctctgccgg aatctttgtg
     1921 aagaatggcc ctgcaatatc cactattagc aaagacatct ttggcgagtg gaacgttatc
     1981 agggataagt ggaatgccga gtacgatgat attcatctca agaagaaagc cgtggttaca
     2041 gagaaatacg aggatgatag acgcaagagc tttaagaaga ttggtagctt ctctctcgaa
     2101 cagctgcagg agtacgccga cgctgacctg tcagtcgtgg agaaactcaa ggagatcata
     2161 atccagaagg tggatgaaat ctacaaagtg tatggaagct ctgagaaact cttcgatgca
     2221 gactttgttc tggagaagag tctgaagaag aacgacgcag tggttgctat catgaaggac
     2281 ctgctggatt ctgttaagtc tttcgagaat tacattaagg cattctttgg tgaagggaag
     2341 gagacaaata gggacgagag cttctatggc gactttgttc tggcctacga catcctcctc
     2401 aaggttgacc acatctatga cgctatacgg aattacgtta cccagaagcc ctatagcaaa
     2461 gacaagttca agctgtattt ccagaatcca cagtttatgg gtgggtggga taaagacaaa
     2521 gaaacagatt acagggccac tatcctgcgg tacggcagca aatactatct ggctatcatg
     2581 gataagaagt acgccaaatg cctccagaag atcgacaagg acgacgtgaa cggtaactac
     2641 gagaagatca attacaagct cctgccagga cctaacaaga tgctgcccaa ggtgttcttc
     2701 tccaagaaat ggatggccta ctataaccca agcgaggaca ttcagaagat atacaagaat
     2761 gggacattca agaagggcga tatgttcaac ctcaacgact gccacaagct gattgatttc
     2821 ttcaaggata gcatttctcg ctatcccaag tggtctaatg catacgattt caacttcagc
     2881 gagactgaga agtacaaaga catcgctggc ttctaccggg aggtggaaga gcaaggctat
     2941 aaggtgtcat tcgaatccgc ttctaagaag gaagtggata agctcgtgga agagggtaag
     3001 ctgtacatgt tccagatata caacaaagac ttcagcgata agagccacgg cactccaaac
     3061 ctccatacta tgtatttcaa gctgctgttt gacgagaaca accacggaca gattaggctg
     3121 tcaggaggcg cagaactctt catgcgcaga gcttcactga agaaggagga actcgttgtc
     3181 cacccagcca atagccctat agccaataag aatccagaca atcctaagaa aaccactact
     3241 ctgtcttacg atgtgtataa ggataagaga ttctctgaag atcagtacga actgcacata
     3301 cccattgcca ttaacaagtg ccctaagaac atcttcaaga ttaacacaga ggttagagtg
     3361 ctcctgaaac acgacgataa cccttatgtt ataggcattg atcgcggaga gagaaacctg
     3421 ctgtacatcg tcgtggtgga cggcaaaggc aacatcgtgg aacagtacag tctcaatgaa
     3481 atcattaaca atttcaacgg aatccgcatt aagaccgact accattctct cctcgacaag
     3541 aaggagaaag aaaggttcga agcaagacag aattggacaa gtatagagaa tatcaaagaa
     3601 ctgaaggctg ggtacatctc tcaggttgtg cacaagatat gtgagctggt ggagaagtac
     3661 gacgctgtta tcgccctcga ggacctgaat agcggcttca agaactccag ggtgaaggtg
     3721 gagaagcagg tgtatcagaa gttcgagaag atgctgatcg acaagctcaa ctatatggtg
     3781 gacaagaaat ccaatccttg cgctactggt ggagccctga agggctatca aatcaccaat
     3841 aagttcgaat ctttcaagtc tatgagcacc cagaatggct tcatcttcta catacccgca
     3901 tggctgacat ccaagattga tccctctacc ggatttgtta atctgctcaa gactaagtac
     3961 acctctattg ctgactcaaa gaagttcata tcatcatttg accgcatcat gtacgtgcca
     4021 gaagaggacc tgttcgagtt tgccctggat tacaagaatt tctctcggac tgacgccgac
     4081 tacatcaaga agtggaagct ctactcttat ggtaatcgga ttcgcatatt ccgcaatccc
     4141 aagaagaata acgtgttcga ttgggaggaa gtttgcctca ccagcgctta caaggagctg
     4201 ttcaataagt atgggattaa ctaccagcag ggcgacataa gagccctgct gtgcgaacaa
     4261 tctgataagg cattctattc ctctttcatg gcactgatgt cactgatgct gcaaatgcgc
     4321 aattccatca ccggaagaac agacgtggac tttctgatct ctcctgtcaa gaactcagat
     4381 ggcatcttct acgattcccg caactatgaa gcacaggaga atgctatcct gcctaagaat
     4441 gccgatgcaa atggagccta taacatcgcc agaaaggtcc tctgggccat aggacaattc
     4501 aagaaagctg aagatgagaa gctggacaag gtgaagatcg ccatttcaaa caaagagtgg
     4561 ctcgaatatg ctcagacctc agtgaagcat ggatcaccca agaagaaacg gaaagtgtga
     4621 gggcccttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc
     4681 ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct
     4741 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     4801 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     4861 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     4921 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     4981 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5041 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5101 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc
     5161 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5221 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5281 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5341 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag
     5401 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     5461 gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa gcatgcatct caattagtca
     5521 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5581 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5641 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5701 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5761 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     5821 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     5881 aaaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
     5941 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
     6001 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac
     6061 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
     6121 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgtcgacag gtgcttctcg
     6181 atctgcatcc tgggatcaaa gcgatagtga aggacagtga tggacagccg acggcagttg
     6241 ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaagcactt cgtggccgag
     6301 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     6361 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     6421 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     6481 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     6541 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     6601 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     6661 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6721 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6781 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6841 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6901 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6961 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7021 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7081 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7141 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7201 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7261 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7321 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7381 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7441 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7501 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7561 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7621 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7681 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7741 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7801 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7861 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7921 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7981 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8041 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8101 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8161 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8221 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8281 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8341 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8401 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8461 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8521 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8581 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8641 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8701 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8761 cctgacgtc
//
