LOCUS       Exported                6276 bp ds-DNA     circular SYN 26-MAR-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6276)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Mar 27, 2019 from SnapGene Viewer 4.3.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6276
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(236..864)
                     /direction=LEFT
                     /label=ColE1 origin
     CDS             complement(1016..1675)
                     /label=AmpR
     promoter        complement(1915..1943)
                     /label=Amp prom
     rep_origin      2014..2454
                     /direction=RIGHT
                     /label=F1 ori
     CDS             complement(2461..2529)
                     /label=LacZ alpha
     primer_bind     2599..2617
                     /label=M13-fwd
     primer_bind     2627..2646
                     /label=T7
     misc_feature    2681..2750
                     /label=attB
     misc_feature    2778..2877
                     /label=SA
     misc_feature    2878
                     /label=Ph2-5'
     CDS             2879..2917
                     /codon_start=1
                     /label=(GGS)4-BHI linker Phase2
                     /translation="SGGSGGSGGSGGS"
     CDS             2918..2971
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2972..4921
                     /codon_start=1
                     /label=GeneSwitch
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLEFPGVDQKKFN
                     KVRVVRALDAVALPQPVGVPNESQALSQRFTFSPGQDIQLIPPLINLLMSIEPDVIYAG
                     HDNTKPDTSSSLLTSLNQLGERQLLSVVKWSKSLPGFRNLHIDDQITLIQYSWMSLMVF
                     GLGWRSYKHVSGQMLYFAPDLILNEQRMKESSFYSLCLTMWQIPQEFVKLQVSQEEFLC
                     MKVLLLLNTIPLEGLRSQTQFEEMRSSYIRELIKAIGLRQKGVVSSSQRFYQLTKLLDN
                     LHDLVKQLHLYCLNTFIQSRALSVEFPEMMSEVIAGSTRYQAEFQYLPDTDDRHRIEEK
                     RKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINY
                     DEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQ
                     AVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLL
                     NQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSI
                     ADMDFSALLSQISS"
     3'UTR           4922..5792
                     /label=hsp70 3'UTR
     CDS             5800..5829
                     /codon_start=1
                     /label=(GGS)4 linker Phase2
                     /translation="GGSGGSGGSG"
     misc_feature    5830..5831
                     /label=Ph2-3'
     misc_feature    5832..5952
                     /label=SD
     misc_feature    complement(5980..6049)
                     /label=attB
     primer_bind     complement(6087..6106)
                     /label=T3
     primer_bind     complement(6124..6142)
                     /label=M13-rev
     misc_binding    complement(6150..6172)
                     /label=LacO
     promoter        complement(6177..6206)
                     /label=lac
ORIGIN
        1 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
       61 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      121 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      181 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      241 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      301 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      361 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      421 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      481 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      541 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      601 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      661 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
      721 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      781 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      841 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      901 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      961 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1021 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1081 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1141 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1201 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1261 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1321 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1381 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1441 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1501 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1561 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1621 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1681 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1741 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1801 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1861 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1921 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1981 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     2041 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     2101 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     2161 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     2221 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     2281 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     2341 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     2401 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     2461 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     2521 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     2581 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
     2641 atagggcgaa ttggagctcg ctagcgatgt aggtcacggt ctcgaagccg cggtgcgggt
     2701 gccagggcgt gcccttgggc tccccgggcg cgtactccac ctcacccatc tggtccatca
     2761 tgatggtcga ctctagaagt cgatccaaca tggcgacttg tcccatcccc ggcatgttta
     2821 aatatactaa ttattcttga actaatttta atcaaccgat ttatctctct tccgcaggtc
     2881 gggaggttcc ggtggaagcg gaggtagcgg cggatccgaa ggacgcggaa gcctgttgac
     2941 gtgcggagat gtggaagaga atccaggacc gatgaagcta ctgtcttcta tcgaacaagc
     3001 atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga agtgcgccaa
     3061 gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt ctccgctgac
     3121 tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc tatttctact
     3181 gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac aggatataaa
     3241 agcattgtta gaattcccgg gtgtcgacca gaaaaagttc aataaagtca gagttgtgag
     3301 agcactggat gctgttgctc tcccacagcc agtgggcgtt ccaaatgaaa gccaagccct
     3361 aagccagaga ttcacttttt caccaggtca agacatacag ttgattccac cactgatcaa
     3421 cctgttaatg agcattgaac cagatgtgat ctatgcagga catgacaaca caaaacctga
     3481 cacctccagt tctttgctga caagtcttaa tcaactaggc gagaggcaac ttctttcagt
     3541 agtcaagtgg tctaaatcat tgccaggttt tcgaaactta catattgatg accagataac
     3601 tctcattcag tattcttgga tgagcttaat ggtgtttggt ctaggatgga gatcctacaa
     3661 acacgtcagt gggcagatgc tgtattttgc acctgatcta atactaaatg aacagcggat
     3721 gaaagaatca tcattctatt cattatgcct taccatgtgg cagatcccac aggagtttgt
     3781 caagcttcaa gttagccaag aagagttcct ctgtatgaaa gtattgttac ttcttaatac
     3841 aattcctttg gaagggctac gaagtcaaac ccagtttgag gagatgaggt caagctacat
     3901 tagagagctc atcaaggcaa ttggtttgag gcaaaaagga gttgtgtcga gctcacagcg
     3961 tttctatcaa cttacaaaac ttcttgataa cttgcatgat cttgtcaaac aacttcatct
     4021 gtactgcttg aatacattta tccagtcccg ggcactgagt gttgaatttc cagaaatgat
     4081 gtctgaagtt attgctgggt cgacgagata tcaagcagaa ttccagtacc tgccagatac
     4141 agacgatcgt caccggattg aggagaaacg taaaaggaca tatgagacct tcaagagcat
     4201 catgaagaag agtcctttca gcggacccac cgacccccgg cctccacctc gacgcattgc
     4261 tgtgccttcc cgcagctcag cttctgtccc caagccagca ccccagccct atccctttac
     4321 gtcatccctg agcaccatca actatgatga gtttcccacc atggtgtttc cttctgggca
     4381 gatcagccag gcctcggcct tggccccggc ccctccccaa gtcctgcccc aggctccagc
     4441 ccctgcccct gctccagcca tggtatcagc tctggcccag gccccagccc ctgtcccagt
     4501 cctagcccca ggccctcctc aggctgtggc cccacctgcc cccaagccca cccaggctgg
     4561 ggaaggaacg ctgtcagagg ccctgctgca gctgcagttt gatgatgaag acctgggggc
     4621 cttgcttggc aacagcacag acccagctgt gttcacagac ctggcatccg tcgacaactc
     4681 cgagtttcag cagctgctga accagggcat acctgtggcc ccccacacaa ctgagcccat
     4741 gctgatggag taccctgagg ctataactcg cctagtgaca ggggcccaga ggccccccga
     4801 cccagctcct gctccactgg gggccccggg gctccccaat ggcctccttt caggagatga
     4861 agacttctcc tccattgcgg acatggactt ctcagccctg ctgagtcaga tcagctccta
     4921 agggggtgac gcctgccctc cccagagcac tgggttgcag gggattgaag ccctccaaaa
     4981 gcacttacgg attctggtgg ggtgtgttcc aactgccccc aactttgtgg atgtcttcct
     5041 tggagggggg agccatattt tattctttta ttgtcagtat ctgtatctct ctctcttttt
     5101 ggaggtgctt aagcagaagc attaacttct ctggaaaggg gggagctggg gaaactcaaa
     5161 cttttcccct gtcctgatgg tcagctccct tctctgtagg gaactctggg gtcccccatc
     5221 cccatcctcc agcttctggt actctcctag agacagaagc aggctggagg taaggccttt
     5281 gagcccacaa agccttatca agtgtcttcc atcatggatt cattacagct taatcaaaat
     5341 aacgccccag ataccagccc ctgtatggca ctggcattgt ccctgtgcct aacaccagcg
     5401 tttgaggggc tggccttcct gccctacaga ggtctctgcc ggctctttcc ttgctcaacc
     5461 atggctgaag gaaaccagtg caacagcact ggctctctcc aggatccgac gcgtcgagct
     5521 tatcgatacc gtcgactaaa gccaaataga aattattcag ttctggctta agtttttaaa
     5581 agtgatatta tttatttggt tgtaaccaac caaaagaatg taaataacta atacataatt
     5641 atgttagttt taagttagca acaaattgat tttagctata ttagctactt ggttaataaa
     5701 tagaatatat ttatttaaag ataattcgtt tttattgtca gggagtgagt ttgcttaaaa
     5761 actcgtttag atccactagt tctagagcgg ccggatcccg gaggtagcgg tggaagcgga
     5821 ggttccggca ggtaagttat tgaacaatgg catcaaatgc cttcatcatc actacccttt
     5881 agcccttaag accccacaat gaccttaccc actcagagaa aaaagtaaat atgaaagccc
     5941 atttgaactt ctaagcttgt cgaccatcat gatggaccag atgggtgagg tggagtacgc
     6001 gcccggggag cccaagggca cgccctggca cccgcaccgc ggcttcgaga ccgtgaccta
     6061 catcatgcat ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     6121 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6181 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6241 taattgcgtt gcgctcactg cccgctttcc agtcgg
//
