LOCUS       Exported                6493 bp ds-DNA     circular SYN 28-NOV-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6493)
  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..6493
                     /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..1431
                     /label=sali arm
     primer_bind     complement(1412..1449)
                     /label=Primer 2
     CDS             1438..1833
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             1843..1899
                     /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             1900..1923
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1933..2253
                     /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             2260..2286
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2287..3000
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     3019..3056
                     /label=Primer 3
     misc_feature    3037..3904
                     /label=kpni arm
     CDS             3037..3327
                     /codon_start=1
                     /translation="AAQAAAAAQAAAAAQAAAAQAAQAEAAESWYLALLGFAEHFRTSS
                     PPKIRLCVHCLQAVFPFKPPQRIEARTHLQLGSVLYHHTKNSEQARSHLEKA"
     primer_bind     complement(3887..3940)
                     /label=Primer 4
     promoter        complement(3996..4014)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4035..4051)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4059..4075
                     /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(4083..4113)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4128..4149
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4437..5025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5196..6056)
                     /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(6057..6161)
                     /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 cctctgaggc ctatccagcc tctgtggtga aacctctgcc
      241 ttcctataga cagcatctac tcatatggaa catcttgtac tcaagaaaga cttttcatgg
      301 agccagagag atggctcaga ggttaagagt gcttgtcact cttccagaga atccaagcaa
      361 acatccaggt ggcctactat aactccagct ccaggtactc acacacaaag acataaagac
      421 aaatggattt ttaaaaactg ttaagccttc cctgaccttc aatatattag cagcatccaa
      481 cctcacgtca cgagaactag ttctttgcaa aatttgcttc caggtgcact gacacgcaag
      541 cttagcacag cgtttggcac ccagcaagtg tggaagcata tggtgaatgg gcaaatgcat
      601 ttcatcctag gggcacaggg agagctaaac ccgtaaccgt gggaaaccgc cctggctgac
      661 ccagtgcagt tcggaaacgc acatttccca caaggcagga cccaagccaa aggggaccgg
      721 aggggcagga gggatgtccc aggctgtccg ctgtggatgc caccggcccg agagcagttt
      781 attgggaaag cccatgcggt tcccacccgg cgcccaggcc tttgagggag accaggagcg
      841 ctcctcagcc acgggcgggg gttggggggc gaacccgagt ctggggtctc ggcagaccag
      901 agaagggcac acgcaacgtt actgtaagtt tccctggggc tgaggcgaga agtccgcgtg
      961 tcaggtccgt gacagaaagt agggtgaaca ggagacgtgc agtcgcgaga acccaggcgg
     1021 gcggcgaagg gaagtcgggg gatgttacct gcaacatccc ggttgtccat cttgagactc
     1081 atccaatccc acaatgctcc ggcgccacct gagaagaggg aggctctgtt atagtgcgct
     1141 tggtctgatg gtaaatgtag tccttctccg acaatctcgt ccaaccccct tctaggacac
     1201 cgggcgtctt ctccacagaa acgccggcgt cagttcacga acgtgtatta ctctcagtta
     1261 cgatgatgca gaggtatcat cctctcctca gagtcttcct cagtcggagc cacttcgaac
     1321 acacaccata agacccagaa gtccttgcgt agcagttctc gcgagaattg cgggtcgcca
     1381 ggccccgccc ctcaaggtgc ttccgcctct ctgtggaggc ggcttgttgt tgtcgacatg
     1441 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
     1501 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
     1561 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
     1621 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
     1681 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
     1741 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
     1801 gaattgctgc cctctggtta tgtgtgggag ggcggttccg gagccacgaa cttctctctg
     1861 ttaaagcaag caggagacgt ggaagaaaac cccggtcccg attacaagga tgacgatgac
     1921 aagggttccg gcggcgtgca ggtggagact atctccccag gagacgggcg caccttcccc
     1981 aagcgcggcc agacctgcgt ggtgcactac accgggatgc ttgaagatgg aaagaaagtt
     2041 gattcctccc gggacagaaa caagcccttt aagtttatgc taggcaagca ggaggtgatc
     2101 cgaggctggg aagaaggggt tgcccagatg agtgtgggtc agagagccaa actgactata
     2161 tctccagatt atgcctatgg tgccactggg cacccaggca tcatcccacc acatgccact
     2221 ctcgtcttcg atgtggagct tctaaaactg gaagtcgatt atccgtacga cgtaccagac
     2281 tacgcagtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2341 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     2401 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2461 accctcgtga ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg
     2521 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2581 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2641 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2701 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     2761 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     2821 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2881 cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     2941 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3001 ggtggaggtg gatctggtgg cggcggttca ggtaccgcgg cacaggcggc ggctgcggcg
     3061 caggcggcgg ctgcagctca ggctgccgcc gcacaggccg cacaggccga ggcagccgag
     3121 tcgtggtatc tggctctcct gggtttcgcc gagcacttcc gaacatctag cccgcccaag
     3181 atccgcttgt gcgttcattg cctgcaggcc gtgttcccct tcaagccgcc tcagcggatc
     3241 gaggcccgga cgcacctgca gctcggctcg gtgctttacc accacacgaa gaacagcgag
     3301 caggcgcgca gccacctgga gaaggcggtg agtgtgcggg ccaggcgggc gggccgcggg
     3361 taccggggac tcgatcctct ccgcctcgtc gggcttggga cttccgaagg cctcgccctc
     3421 acctgcggtg acagacgcgg ctccgccacg ccctctcacc cgggaaccgg gaaagttgtc
     3481 cctcttcagc tacttggtgc ttgcatcggg gaccctccta gtcgggtggc cgcgggaaca
     3541 ggacccagag cgtcagccta gaagcaccta cctaaggaag agatgcgagc tccctgacgt
     3601 gtgaaagcga ggtcgcagga aatctgaaag tccaggactt ctgacagtca ggtgtctagg
     3661 tctggagtcc cagatccttg tggcatggat attggggaag gtgaggcctt cagcctcttg
     3721 ttgaacgttg ttgccttgtg aggacctgaa gtgcagaaag tagcttctag agtcacttgc
     3781 acctcggggg tctggaggtc agtctcactg ttggtggctt cttctcccac cacgaaatgg
     3841 tccatctttg aaagagtccc cactccgcct ccaacttgat ttgtgacgtc ttggtgcttc
     3901 acagggtacc tgggcccccc ctcgaggtcg atcgacggta tcgataagct tgatctagag
     3961 cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattccgagc
     4021 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4081 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4141 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4201 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4261 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4321 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4381 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4441 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4501 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4561 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4621 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4681 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4741 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4801 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4861 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4921 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4981 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5041 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5101 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5161 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5221 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5281 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5341 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5401 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5461 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5521 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5581 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5641 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5701 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5761 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5821 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5881 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5941 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6001 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6061 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6121 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6181 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     6241 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     6301 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     6361 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     6421 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     6481 taccgcatca ggc
//
