LOCUS       Exported                6577 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 6577)
  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..6577
                     /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..1487
                     /label=sali arm
     primer_bind     complement(1468..1505)
                     /label=Primer 2
     primer_bind     complement(1468..1496)
                     /label=Neo 3'
     CDS             1494..1889
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             1899..1955
                     /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             1956..1979
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1989..2309
                     /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             2316..2342
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2343..3056
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     3075..3112
                     /label=Primer 3
     misc_feature    3093..3988
                     /label=kpni arm
     CDS             3093..3314
                     /codon_start=1
                     /translation="STPTDPAAMPHPGPSPGPGPSPGPILGPSPGPGPSPGSVHSMMGP
                     SPGPPSVSHPLSTMGSADFPQEGMHQLHK"
     primer_bind     complement(3971..4024)
                     /label=Primer 4
     promoter        complement(4080..4098)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4119..4135)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4143..4159
                     /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(4167..4197)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4212..4233
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4521..5109)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5280..6140)
                     /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(6141..6245)
                     /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 cgccaggaag cttagaggct gggtgatggg gggagtgttc
      241 agttaagtct ggagaaacag aacgcccatt cgcagacaca gacagctaca ccctggcatc
      301 cttgaccaga ggcctagcac gaatcatgag tcttagttgt ccaggttttg gttctaatac
      361 ttaacctagg aggaggaccc cagcgttgtc cattttgata acttgaggtc cagtagctgt
      421 ggtgggtagg tgaggcaggc agctgtggtt ggaattctgc tgttgtttta catgtatgtc
      481 tcatctgact ctaagaagta tcaagacaag aacttatcta ataattgtgt ctgaatctat
      541 ttgagcctat attgtatgat ccctgtagct tgcttgctct ctctctctct ctctctctcc
      601 ctctctctct ctcccctttc tctctctccc tccctctccc cctctccttc cctctctccc
      661 tctccccctt cccctcccct tttcccctcc ccttccctct cccctcccac atttcctctc
      721 cctctctctg ttttgttgtt gttttgtttt tgaggcaggg tttctctgtg tagtgctggt
      781 tgtcctggaa cttgctctct ccaccaggct ggcttggacc tcacagagct cctcctgctt
      841 ctgcctccca agtgctagga ggcacctatg tgccaccctg tgtgttattt ttcttccttt
      901 tctataattg agacttctct ttttattatc aattgatgga cttatttttc ccccttttag
      961 ctatctagaa atcttggtga agttttgatt gcctgttggc ttgtgaaaat gagtcttggt
     1021 tggatactta tttgcaaatg gttttccagt ccagtggttt gggggagtgg gggcttaatg
     1081 acttctgagg tgtctaagcc agcctataac agagcgtatc tgttgtgttt agtagtaaat
     1141 ttcagacttg tttagattct gcttatgtgt taaatttcag ttgcttcagt gagtgtctct
     1201 atatccaggg tgatttatca tttgggggga atcacttaaa gtatgtgttt tttaaaacaa
     1261 gtggtcttca taaaatctga gcccagggtt atatatcaca ttcattcaaa catctggaag
     1321 aggccaggga gttctgtttg atgttagtta cagaaatggc accatcagat tttaacaact
     1381 gttttatttg gaccttaaca tcatgtttaa aacatttctt tctttctttt tcttcctgct
     1441 ctccaactta gtgtcactct gctaactggc agagccagga gacctaggtc gacatggcca
     1501 agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca
     1561 tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct
     1621 tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg
     1681 gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga
     1741 acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg
     1801 ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat
     1861 tgctgccctc tggttatgtg tgggagggcg gttccggagc cacgaacttc tctctgttaa
     1921 agcaagcagg agacgtggaa gaaaaccccg gtcccgatta caaggatgac gatgacaagg
     1981 gttccggcgg cgtgcaggtg gagactatct ccccaggaga cgggcgcacc ttccccaagc
     2041 gcggccagac ctgcgtggtg cactacaccg ggatgcttga agatggaaag aaagttgatt
     2101 cctcccggga cagaaacaag ccctttaagt ttatgctagg caagcaggag gtgatccgag
     2161 gctgggaaga aggggttgcc cagatgagtg tgggtcagag agccaaactg actatatctc
     2221 cagattatgc ctatggtgcc actgggcacc caggcatcat cccaccacat gccactctcg
     2281 tcttcgatgt ggagcttcta aaactggaag tcgattatcc gtacgacgta ccagactacg
     2341 cagtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2401 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2461 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2521 tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc
     2581 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2641 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2701 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2761 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     2821 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2881 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2941 acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3001 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggtg
     3061 gaggtggatc tggtggcggc ggttcaggta cctccacacc cacagaccca gcagcaatgc
     3121 cccatcctgg gccctccccg gggcctggac cctctcctgg accaattctg gggcctagtc
     3181 caggaccagg accatcccca ggttctgtgc acagcatgat gggtcctagt cccggacctc
     3241 ccagcgtctc acatcctctg tcaacgatgg gctctgcaga cttcccacag gaaggcatgc
     3301 accaattaca taaggtaaga gtctgtgccc tctcccccac tttacccgtc ttctcaggca
     3361 ttctgatgac tctgagcttt ctcctgctag taacataaga atctagctag aaatttgtgt
     3421 taacttgcat gttattaaaa acattcctgt aatcccagca cttgagagac tgaagcaaaa
     3481 ggatcatgca ttaaagacca atctggatct tataaaaacc caaaccaaac cacaaccaag
     3541 cccaatataa agcaaaaacc atccccaaac tggggtaaaa cacttctact gctgtatttt
     3601 ttactgttac ggatggccca ggaatcgaga agttgtcttt tcaaacacaa aggtccgcca
     3661 tgcaatttgg acaggtttgg ggattggtgg gttaaaggta gtttgtcttg gtgaacgtgt
     3721 atctcagtaa gggagacttc cttccctcac ctcccatgta ctttatatca ctcacaaatc
     3781 aatatattag aaattagtcc tccctacaaa tgttgaaatt tatttgatga aactagatct
     3841 gtaagagtga aggtgaaagt tgccttctga aggttttgca gggcctggcg agagagagag
     3901 agagagagag agagagagag agagagagag agagagagat cagcacctaa gagcatgtga
     3961 agcactacca ttgggcagct cacactgggg tacctgggcc ccccctcgag gtcgatcgac
     4021 ggtatcgata agcttgatct agagcggccg ccaccgcggt ggagctccag cttttgttcc
     4081 ctttagtgag ggttaattcc gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4141 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4201 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4261 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4321 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4381 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4441 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4501 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4561 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4621 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4681 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4741 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4801 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4861 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4921 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     4981 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5041 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5101 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5161 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5221 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5281 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5341 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5401 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5461 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5521 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5581 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5641 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5701 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5761 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5821 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5881 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5941 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6001 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6061 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6121 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6181 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6241 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     6301 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     6361 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     6421 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     6481 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     6541 ccgcacagat gcgtaaggag aaaataccgc atcaggc
//
