LOCUS       Exported                4365 bp ds-DNA     circular SYN 11-DEC-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4365)
  AUTHORS   Molecular Function Laboratory
  TITLE     Direct Submission
  JOURNAL   Exported Dec 11, 2018 from SnapGene Viewer 4.2.9
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4365
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    701..706
                     /label=Kozak seq
     CDS             707..1102
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     960..982
                     /label=(1242) BSD seq1
     CDS             1115..1171
                     /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             1178..1382
                     /codon_start=1
                     /label=Linker-mAID-Clover
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             1178..1381
                     /codon_start=1
                     /label=mAID
                     /note="note=""direct strand 
                     translation""translation=""KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNV
                     MVSCQKSSGGPEAAAFVKVSMDGAPYLRKIDLRMYK"""
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             complement(1386..2083)
                     /codon_start=1
                     /label=ORF frame 1
                     /translation="MCNPGGGHELQQDHVIALLVGVFAQLGLMAQVVVVGQQHGAVADG
                     GVLLVVVGELHAAVLNVVADLEVSLDAVLLLVGRDIDVVAVEVVLQLVPQDVAVLLEVD
                     ALQLDAVHQGVALELHLGAGLVGTVVLERDGALLDVAFGHGGLEEVVLLHVVGVAAEAG
                     HAVAEGGHEGGPGHGQLAGGADELQGQLAVGGIALALAADAELVAVYVAVQLDQDGHHP
                     GEQLLALAHH"
     CDS             1388..2104
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             1388..2097
                     /codon_start=1
                     /label=ORF frame 2
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDE"
     primer_bind     1388..1408
                     /label=(1019)_CCC_FP_F
     gene            1391..2097
                     /gene="EGFP (variant)"
                     /label=EGFP (variant)
     gene            1391..2097
                     /gene="ECFP (variant)"
                     /label=ECFP (variant)
     misc_feature    complement(1433..1454)
                     /label=EGFP_N_primer
     primer_bind     1935..1954
                     /label=(345) GFP seq 1
     primer_bind     complement(1991..2023)
                     /label=(1018)_mClover(A206K)_SDM_R
     primer_bind     1991..2023
                     /label=(1017)_mClover(A206K)_SDM_F
     misc_feature    2006..2008
                     /label=A206K
     misc_feature    2041..2062
                     /label=EGFP_C_primer
     primer_bind     complement(2085..2104)
                     /label=(1020)_CCC_mClo_TAA_NotI_R
     CDS             2105..2122
                     /codon_start=1
                     /label=Linker
                     /translation="GAGAGA"
     CDS             2111..2122
                     /codon_start=1
                     /label=Linker-mAID-Clover
                     /translation="GAGA"
     misc_feature    2111..2120
                     /label=(965)_pAID_Linker_5'_BamHI_SacI
     promoter        complement(2177..2195)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2216..2232)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2240..2256
                     /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(2264..2294)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2309..2330
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2618..3206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3377..4237)
                     /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(4238..4342)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gccaccatgg ccaagccttt
      721 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
      781 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
      841 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
      901 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
      961 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
     1021 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
     1081 ctctggttat gtgtgggagg gctcaggaag cggagctact aacttcagcc tgctgaagca
     1141 ggctggagac gtggaggaga accctggacc tagatctaag gagaagagtg cttgtcctaa
     1201 agatccagcc aaacctccgg ccaaggcaca agttgtggga tggccaccgg tgagatcata
     1261 ccggaagaac gtgatggttt cctgccaaaa atcaagcggt ggcccggagg cggcggcgtt
     1321 cgtgaaggta tcaatggacg gagcaccgta cttgaggaaa atcgatttga ggatgtataa
     1381 agctagcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1441 gctggacggc gacgtaaacg gccacaagtt cagcgtccgc ggcgagggcg agggcgatgc
     1501 caccaacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1561 gcccaccctc gtgaccacct tcggctacgg cgtggcctgc ttcagccgct accccgacca
     1621 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1681 catctctttc aaggacgacg gtacctacaa gacccgcgcc gaggtgaagt tcgagggcga
     1741 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1801 ggggcacaag ctggagtaca acttcaacag ccacaacgtc tatatcacgg ccgacaagca
     1861 gaagaacggc atcaaggcta acttcaagat ccgccacaac gttgaggacg gcagcgtgca
     1921 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1981 caaccactac ctgagccatc agtccaagct gagcaaagac cccaacgaga agcgcgatca
     2041 catggtcctg ctggagttcg tgaccgccgc cgggattaca catggcatgg acgagctgta
     2101 caagggcgct ggtgcaggcg ccggatccac tagttctaga gcggccgcca ccgcggtgga
     2161 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     2221 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2281 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2341 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2401 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2461 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2521 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2581 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2641 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2701 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2761 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2821 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2881 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2941 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3001 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     3061 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3121 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3181 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3241 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3301 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3361 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3421 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3481 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3541 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3601 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3661 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3721 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3781 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3841 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3901 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3961 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4021 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4081 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4141 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4201 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4261 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4321 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
