LOCUS       Exported                6780 bp ds-DNA     circular SYN 01-OCT-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6780)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Nov 24, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6780
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     142..158
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     182..219
                     /label=Primer 1
     misc_feature    202..1391
                     /label=SalI arm
     primer_bind     complement(1374..1411)
                     /label=Primer 2
     CDS             1398..1793
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             1803..1859
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1860..1883
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1893..2213
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2220..2246
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2247..2960
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     2977..3014
                     /label=Primer 3
     misc_feature    2997..4192
                     /label=KpnI arm
     CDS             2997..3086
                     /codon_start=1
                     /translation="HGGGPPSGDSACPLRTIKRVQFGVLSPDEL"
     primer_bind     complement(4175..4227)
                     /label=Primer 4
     promoter        complement(4283..4301)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4322..4338)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4346..4362
                     /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(4370..4400)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4415..4436
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4724..5312)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5483..6343)
                     /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(6344..6448)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      121 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc cactagtggc cgcggcggcc
      181 gcagcttgcc tgcaggtcga caagtcgtct gaaaggtgta gggtttcctt gaccccgaat
      241 tccccaagac atccctattc tgccccaccg ctaatctgct tttctcagat tgtcacggtc
      301 gcgtgatctg cggctcctgg gtagcagcca ttctactttc agaataggcc tccgggctcg
      361 ggaaaagctt gggctgaaca tcgagcgcca gggctccgta aagctattag agcacaggcg
      421 gtgccccaac gtcctggggc ctctccacta ataacggcta cttccaattg attggacgcg
      481 ccatcttgcc tgccttatgc atattcagcg gtgaactgaa tattcatgaa cgaggcccgt
      541 cccgtccctc cctccttccc cccacccccg gaacccgctc cggaggaccc gaagggcccc
      601 gccttcatta ccgatgcgta ggacaaacca ttttcccgat gtgtgtgggg ggatactaat
      661 gagagacttt agctgaaaat gagcctgaac tccgaagctg agtaaaaatg gcctaacttt
      721 atcctccgtt ctgtaagtcc tcggtttgag tgcacgggaa acccgaaagg aggacgacag
      781 gaccaggaac attctcctcc tcctgtcgcg tcagaaagaa cacccaacca gggagccgga
      841 gccctagcgt caacaactcc gccgcgcgcg ctccgtgtag gccggtgcgg gcggccccgt
      901 agcgcaaggg agggcgggaa aggaaggggc gggacacaag ggcgaatcta taaagggcgt
      961 cactcagcca gttctctcct cagaagcgcc gagagcgcga ccgggacggt tggagaagaa
     1021 ggtggctccc ggaaggggga gagacaaact gccgtaacct ctgccgttca ggatcccggt
     1081 tacttattta ttcgttaccc cttttcttct tcctccctcc caaaacctcc tcctctcctc
     1141 ccttcttttg tttccttttt gggagccgac gaatctccgg aaagggagaa aaggctttct
     1201 ttcagccctt ttcgttcctc tgccttcccc cccccttccc ccctccccca cctttccctc
     1261 ctccagcctt tccctcccta tcccgggagg tctttcctgg tggccgcccg gacgggttct
     1321 gagcacttag gcggcggtgg cgcaggcttt ttgtagcgag gtttgcgcct gcgcagcgcg
     1381 cctgcctcgc cgtcgacatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa
     1441 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag
     1501 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac
     1561 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt
     1621 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac
     1681 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc
     1741 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggcggttccg
     1801 gagccacgaa cttctctctg ttaaagcaag caggagacgt ggaagaaaac cccggtcccg
     1861 attacaagga tgacgatgac aagggttccg gcggcgtgca ggtggagact atctccccag
     1921 gagacgggcg caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc
     1981 ttgaagatgg aaagaaagtt gattcctccc gggacagaaa caagcccttt aagtttatgc
     2041 taggcaagca ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc
     2101 agagagccaa actgactata tctccagatt atgcctatgg tgccactggg cacccaggca
     2161 tcatcccacc acatgccact ctcgtcttcg atgtggagct tctaaaactg gaagtcgatt
     2221 atccgtacga cgtaccagac tacgcagtga gcaagggcga ggagctgttc accggggtgg
     2281 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     2341 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2401 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg cagtgcttcg
     2461 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2521 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2581 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2641 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2701 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2761 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2821 ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca
     2881 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2941 gcatggacga gctgtacaag ggtggaggtg gatctggtgg cggcggttca ggtacccacg
     3001 ggggtggccc cccctccggg gacagcgcat gcccgctgcg caccatcaag agagtgcagt
     3061 tcggagtcct gagtccggat gaattggtaa gatagtccgc ccgtctctct gctatgcctt
     3121 tggctgatga tcctctggga agatgggtca gtgatgctga tgttgtgcgg gggtccagcc
     3181 ttcaaaagtt aaattatgcc aggacttaaa caagcgaaat gctatgcctg agatgagtgc
     3241 ttgactggac agagaagaaa gatgtttcca tacttgagca ttgctgcggt tcaacgagac
     3301 agggttgaaa gttgtacagg aagtggatcc ccagttttag ttctttcacc aatattgagc
     3361 atcctgctat gtgacagtaa tcagacacta ggaacacagc tgcacaataa atataacaga
     3421 aactctacct tctaaaacac tatgggaaaa acagagaaga gagcttaaga acaaaaagaa
     3481 atgagcaaaa ataagtacct agattaaagc aaaatgagga aatgctgtaa gaagtcactg
     3541 aaattgaatg gtgtcagggt ttggaagaca tacttgccag ggatggtgac attcccaagt
     3601 aatatcagtc atcagagcta tttttttagg tggaggaaac aagtgtcatg ggactgagag
     3661 ggaaataaac ttgtattcta gagactgaaa agccagtgtt ggggcaggca gggttagtgg
     3721 gtgcggagag tgcactgaag aagcacggga tattgtaaat tacgctctag agagtccaaa
     3781 tttctgtttt cagtaggaaa ccactgggta attataaagg tgacagtagt aacttataca
     3841 cacacgcatg tacagaagct ccgtaaagag ctgatgtgaa gtgggtgagg atagacaagg
     3901 gatagctttt ggagacttgg acttcttcct cctgagagga atagagagtg gaattaaaca
     3961 gctgattcta gactgaggca gtcctgttgg ggtaagggct caactatagc agaactcaaa
     4021 agaaaaaaca gaaaaaccta catcagggct ggtaagatgg ctcagcaggt aagagaactg
     4081 actgctcttc cgaaagtcat gagttcaaat cccagcaacc atatggtggc tcacaaccat
     4141 ctgtaatgag atctgatgcc cttttctgga gtgtctgaag tcagctacag tgggtaccgg
     4201 gccccccctc gaggtcgatc gacggtatcg ataagcttga tctagagcgg ccgccaccgc
     4261 ggtggagctc cagcttttgt tccctttagt gagggttaat tccgagcttg gcgtaatcat
     4321 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4381 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4441 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4501 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4561 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4621 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4681 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4741 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4801 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4861 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4921 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4981 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5041 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5101 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5161 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5221 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5281 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5341 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5401 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5461 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5521 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5581 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5641 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5701 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5761 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5821 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5881 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5941 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6001 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6061 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6121 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6181 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6241 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6301 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6361 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6421 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     6481 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     6541 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     6601 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     6661 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     6721 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
//
