LOCUS       Exported                6926 bp ds-DNA     circular SYN 09-DEC-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6926)
  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..6926
                     /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..1461
                     /label=salI arm
     primer_bind     complement(1444..1481)
                     /label=Primer 2
     CDS             1468..1863
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             1873..1929
                     /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             1930..1953
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1963..2283
                     /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"
     primer_bind     2290..2334
                     
     CDS             2290..2316
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2317..3030
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     2893..2912
                     /label=EGFP/EYFP AA 195 5'
     primer_bind     3047..3084
                     /label=Primer 3
     misc_feature    3067..4338
                     /label=KpniI arm
     CDS             3067..3139
                     /codon_start=1
                     /translation="PGPPSAKRPKLSSPALSASASDGT"
     primer_bind     complement(4321..4373)
                     /label=Primer 4
     promoter        complement(4429..4447)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4468..4484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4492..4508
                     /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(4516..4546)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4561..4582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4870..5458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5629..6489)
                     /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(6490..6594)
                     /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 cggcagaggc aagaaacaac agccgccatc ttgtttgtgt
      241 gctagcctgt ggggggggtg gggggaggag gagagagagc ggagagggac cgggagagag
      301 agagtggggc aagcgggacg cccggctgag tgcgaactgc gcgagcaggg agtgaggagg
      361 gtacctggct ggagagcagc ggccgggacc gagacagtgg ttgggggccc cggggcgcaa
      421 aggctgaggc gaccctcagt cccctccttg cccgcacaca cccccaccgc agcccgggcc
      481 ggcgtcgacg cctaaggggg ggaaaccatt acataagccc gcgccccgcg gcgcctctgc
      541 ccgccgtccg aggcggcccc cgagccgagc ccccggcagg cacctggctg ccgggggggt
      601 gagggccagg gaggcggccg gactgatttc tcctccccgg gcttgcgagt ggaggcggag
      661 aagcgcggcg cgcggagagg ccgcgtcggt gggctcggcg tcagcgcggg cttgcgggcg
      721 gacggccctg cttggcccgg cccggccccc ctggcccttc gggccggcga gctccacgct
      781 cggcgtcgtc gctgctgcgc ctctgcttgt ccgctgcgcg atgtgaggcg gccccggcgc
      841 cagtctggct ctcggctcgg ctgagctcag tgtggccatt aggggccggt gcggcagcgg
      901 cggcgcggag cgcagcggca ggaggaggag gagggcggct ctcagggtgg gggcgcaggc
      961 ccgggagggg gcaccgggag gaggtgagtg tctctggtcg cctcctcctc tccccctttt
     1021 tcgcccccgc ctccttgtgg cgatgagaag gaggaggaca gcgccgagga ggaagaagct
     1081 gatggcggcg gcggagctcc gagagacctc ggctgggcag gggctcgccg tggcgggctg
     1141 gggactgcgt ctgtagagct gtgagttccc gagaattcgc tgcagcggac gtgctcgtgg
     1201 gatcagtgtt gctgtaccct cctcccggcc tggtttaggc ccggcccctc gcacttgccc
     1261 ctaccatttc tgttgagtcc gcatccctct ccggccaccg caaccaggcg aagagaaaaa
     1321 ggaacttccc cccacccctg ttgggtggcg gcggagctcc ggagcccacc cttaggagcg
     1381 gggcggggac cccgtttcgg agaagagatt tcctcaggat tctcgttttt ttcctcggtt
     1441 gtatctccga aagaattaaa agtcgacatg gccaagcctt tgtctcaaga agaatccacc
     1501 ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc
     1561 gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt
     1621 actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc
     1681 aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga
     1741 cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt
     1801 gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag
     1861 ggcggttccg gagccacgaa cttctctctg ttaaagcaag caggagacgt ggaagaaaac
     1921 cccggtcccg attacaagga tgacgatgac aagggttccg gcggcgtgca ggtggagact
     1981 atctccccag gagacgggcg caccttcccc aagcgcggcc agacctgcgt ggtgcactac
     2041 accgggatgc ttgaagatgg aaagaaagtt gattcctccc gggacagaaa caagcccttt
     2101 aagtttatgc taggcaagca ggaggtgatc cgaggctggg aagaaggggt tgcccagatg
     2161 agtgtgggtc agagagccaa actgactata tctccagatt atgcctatgg tgccactggg
     2221 cacccaggca tcatcccacc acatgccact ctcgtcttcg atgtggagct tctaaaactg
     2281 gaagtcgatt atccgtacga cgtaccagac tacgcagtga gcaagggcga ggagctgttc
     2341 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2401 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     2461 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
     2521 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2581 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2641 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2701 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2761 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2821 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2881 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     2941 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3001 atcactctcg gcatggacga gctgtacaag ggtggaggtg gatctggtgg cggcggttca
     3061 ggtacccccg ggccgccttc agccaagcgg cctaaactct catctccggc cctctcggcg
     3121 tccgccagcg atggcacagg ttagtctcgg gactcccggc cttccatagt tcccttcgtc
     3181 ttttctgctc gttgcacctt cattccaggg tgcttctgtg tatctcttcc catttccccg
     3241 gccggtgttt aattttaaag gtttttgaat ggactggtcg aagacgtcca gtagcccaac
     3301 cattttcttt gccttctgat acattcattg caacactatg tcatgtcaga tcgttgtgtg
     3361 ttctggaatt aagaggttgg aagtgggagc tgcggggacc tttattgttg gctccatgca
     3421 atttaatttg ggaaatgccg gtacaatcgt gtgtgagggc ttttatgcac ttttactatt
     3481 tttgttactg gtgaatgagg gatttcagct ctttctcccc tgttgcaatg aattcttttt
     3541 ttctctgctt tcccaccttt aatgattttt gctcctgtga ggcagatttt ctgttgagga
     3601 atgggggcgg aagacaaatc ttggtgtctg cgatccaagg ttttaagtgt aactaaagtt
     3661 caacaaaatt tcattcttag gtcccctctc cccatccttg cctcaagagt cagtatggac
     3721 cgcagtttat tttatatcca tttgccatga aaccaaaatg tctcaaactg cgtgttgagg
     3781 tgtagacctg aggcgtttcc tttgcagttt gtcacactct tacttagaag ggaaagaaaa
     3841 aagatgaagc ctttctggaa ttaggtactt agaaatggga gataataagt tgaaagactc
     3901 ccagtcatga acttccaact aaagtgattg tacaagttta aaaatgaaca ggaggttttg
     3961 tgcttgcatg tttgttttgt tttgcttttt ggacctttgc aattccagtt tacaagatga
     4021 aggaattctt ttctccagtc agcccctttg ttaggagacc tgaagggaga ctctcccttg
     4081 tctgttcttt ttcctgctta ctgttagttt agtgttattc tgatgatcac tgacctggtc
     4141 ctctcttgct gtctgacttg tttgagaggc caggataatg aagaagaaga agaagaaaaa
     4201 gaagaagaag aagaagaatg gctagtcatc tgacagtcca ctagatgttc ctccttactg
     4261 caagcatccc tttaactcag agaagaggct ccagatctct ccttctgcca atgagagtgt
     4321 atagtggagt taaacgtagg taccgggccc cccctcgagg tcgatcgacg gtatcgataa
     4381 gcttgatcta gagcggccgc caccgcggtg gagctccagc ttttgttccc tttagtgagg
     4441 gttaattccg agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4501 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4561 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4621 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4681 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4741 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4801 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4861 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4921 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4981 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5041 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5101 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5161 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5221 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5281 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     5341 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5401 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5461 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5521 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5581 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5641 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5701 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5761 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5821 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5881 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5941 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6001 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6061 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6121 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6181 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6241 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6301 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6361 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6421 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6481 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6541 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6601 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     6661 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     6721 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6781 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     6841 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     6901 cgtaaggaga aaataccgca tcaggc
//
