LOCUS       Exported                4465 bp ds-DNA     circular SYN 22-AUG-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Addgene207872_p0720_pRset_mScarlet_BsEpsilon_NanoLuc
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 4465)
  AUTHORS   Tantama Lab
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Aug 22, 2023 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4465
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        20..38
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    84..90
                     /note="RBS"
                     /note="ribosome binding site"
     CDS             100..102
                     /product="start codon"
                     /note="ATG"
     CDS             112..129
                     /product="6xHis affinity tag"
                     /note="6xHis"
     CDS             133..165
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /note="T7 tag (gene 10 leader)"
                     /note="promotes efficient translation in E. coli"
     CDS             169..192
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /note="Xpress(TM) tag"
     CDS             211..866
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /note="mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERS"
     CDS             892..897
                     /codon_start=1
                     /note="NLinker"
                     /translation="ID"
     CDS             898..1293
                     /codon_start=1
                     /note="B.subtilis Epsilon Subunit(H42K)"
                     /translation="MKTVKVNITTPDGPVYDADIEMVSVRAESGDLGILPGHIPTKAPL
                     KIGAVRLKKDGQTEMVAVSGGTVEVRPDHVTINAQAAETAEGIDKERAEAARQRAQERL
                     NSQSDDTDIRRAELALQRALNRLDVAGK"
     CDS             1294..1308
                     /codon_start=1
                     /note="CLinker"
                     /translation="ANEFM"
     CDS             1309..1812
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /note="Nluc"
                     /note="human codon-optimized"
                     /translation="LEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRIVLS
                     GENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVTPNM
                     IDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRLCE
                     RILA"
     terminator      1880..1927
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2024..2479
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2505..2609
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             2610..3470
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3641..4229
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggccacctc gagtaccacc atggtgagca agggcgaggc agtgatcaag
      241 gagttcatgc ggttcaaggt gcacatggag ggctccatga acggccacga gttcgagatc
      301 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
      361 aagggtggcc ccctgccctt ctcctgggac atcctgtccc ctcagttcat gtacggctcc
      421 agggccttca ccaagcaccc cgccgacatc cccgactact ataagcagtc cttccccgag
      481 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgccgtgac cgtgacccag
      541 gacacctccc tggaggacgg caccctgatc tacaaggtga agctccgcgg caccaacttc
      601 cctcctgacg gccccgtaat gcagaagaag acaatgggct gggaagcgtc caccgagcgg
      661 ttgtaccccg aggacggcgt gctgaagggc gacattaaga tggccctgcg cctgaaggac
      721 ggcggccgct acctggcgga cttcaagacc acctacaagg ccaagaagcc cgtgcagatg
      781 cccggcgcct acaacgtcga ccgcaagttg gacatcacct cccacaacga ggactacacc
      841 gtggtggaac agtacgaacg ctccgaagga cgtcattcta caggccgcca tatcgatatg
      901 aaaactgtga aagtgaatat aacaacccct gatgggccag tctacgacgc tgatatcgag
      961 atggtgtccg tgcgggccga gagtggtgat ctcggcatcc tccccggtca cattcccaca
     1021 aaggccccac tgaagatcgg agctgtgcgg ctgaagaagg acggccaaac cgagatggtc
     1081 gcagtctcag gcggcactgt tgaagtgcgg cctgaccacg ttaccattaa tgctcaagcc
     1141 gctgaaacag ccgaaggaat cgacaaagag agagcagaag ccgcaagaca gagggcccag
     1201 gagcggctga actctcaatc cgatgacacc gatattcgcc gggccgagct ggcactgcag
     1261 agggccctga acagactgga cgtggctggg aaggcaaacg aattcatgct ggaggacttt
     1321 gttggggact ggcgacagac agccggctac aacctggacc aagtccttga acagggaggt
     1381 gtgtccagtt tgtttcagaa tctcggggtg tccgtaactc cgatccaaag gattgtcctg
     1441 agcggtgaaa atgggctgaa gatcgacatc catgtcatca tcccgtatga aggtctgagc
     1501 ggcgaccaaa tgggccagat cgaaaaaatt tttaaggtgg tgtaccctgt ggatgatcat
     1561 cactttaagg tgatcctgca ctatggcaca ctggtaatcg acggggttac gccgaacatg
     1621 atcgactatt tcggacggcc gtatgaaggc atcgccgtgt tcgacggcaa aaagatcact
     1681 gtaacaggga ccctgtggaa cggcaacaaa attatcgacg agcgcctgat caaccccgac
     1741 ggctccctgc tgttccgagt aaccatcaac ggagtgaccg gctggcggct gtgcgaacgc
     1801 attctggcgt aagcttgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1861 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1921 ttttttgctg aaaggaggaa ctatatccgg atctggcgta atagcgaaga ggcccgcacc
     1981 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     2041 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2101 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2161 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2221 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2281 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2341 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2401 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     2461 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2521 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2581 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2641 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2701 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2761 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2821 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2881 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2941 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3001 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3061 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3121 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     3181 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3241 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3301 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3361 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3421 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3481 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3541 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3601 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3661 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3721 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3781 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3841 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3901 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3961 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4021 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4081 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4141 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4201 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4261 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4321 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     4381 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     4441 cgcgttggcc gattcattaa tgcag
//
