LOCUS       Exported                4167 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 4167)
  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..4167
                     /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"
     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             1388..1900
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(1876..1924)
                     /label=(1843) Nluc-NarI
     CDS             1901..1924
                     /codon_start=1
                     /label=Linker
                     /translation="GAGAGAGA"
     CDS             1921..1924
                     /codon_start=1
                     /label=Linker-mAID-Clover
                     /translation="A"
     misc_feature    1921..1922
                     /label=(965)_pAID_Linker_5'_BamHI_SacI
     promoter        complement(1979..1997)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2014..2032)
                     /label=(2) M13 reverse primer
     primer_bind     complement(2018..2034)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2042..2058
                     /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(2066..2096)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2111..2132
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2420..3008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3179..4039)
                     /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(4040..4144)
                     /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 gtcttcacac tcgaagattt cgttggggac tggcgacaga cagccggcta
     1441 caacctggac caagtccttg aacagggagg tgtgtccagt ttgtttcaga atctcggggt
     1501 gtccgtaact ccgatccaaa ggattgtcct gagcggtgaa aatgggctga agatcgacat
     1561 ccatgtcatc atcccgtatg aaggtctgag cggcgaccaa atgggccaga tcgaaaaaat
     1621 ttttaaggtg gtgtaccctg tggatgatca tcactttaag gtgatcctgc actatggcac
     1681 actggtaatc gacggggtta cgccgaacat gatcgactat ttcggacggc cgtatgaagg
     1741 catcgccgtg ttcgacggca aaaagatcac tgtaacaggg accctgtgga acggcaacaa
     1801 aattatcgac gagcgcctga tcaaccccga cggctccctg ctgttccgag taaccatcaa
     1861 cggagtgacc ggctggcggc tgtgcgaacg cattctggcg ggcgctggtg caggggccgg
     1921 cgccggatcc actagttcta gagcggccgc caccgcggtg gagctccagc ttttgttccc
     1981 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     2041 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     2101 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     2161 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2221 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2281 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2341 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2401 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2461 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2521 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2581 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2641 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2701 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2761 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2821 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     2881 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2941 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3001 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3061 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3121 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3181 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3241 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3301 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3361 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3421 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3481 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3541 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3601 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3661 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3721 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3781 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3841 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3901 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3961 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4021 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4081 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4141 cgcgcacatt tccccgaaaa gtgccac
//
