LOCUS       Exported                6873 bp ds-DNA     circular SYN 22-AUG-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Addgene207873_p0763_GW1_mScarlet_BsEpsilon_NanoLuc
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6873)
  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..6873
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     regulatory      184..192
                     /regulatory_class="other"
                     /note="Kozak sequence"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             189..844
                     /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             870..875
                     /codon_start=1
                     /note="NLinker"
                     /translation="ID"
     CDS             876..1271
                     /codon_start=1
                     /note="B.subtilis Epsilon Subunit(H42K)"
                     /translation="MKTVKVNITTPDGPVYDADIEMVSVRAESGDLGILPGHIPTKAPL
                     KIGAVRLKKDGQTEMVAVSGGTVEVRPDHVTINAQAAETAEGIDKERAEAARQRAQERL
                     NSQSDDTDIRRAELALQRALNRLDVAGK"
     CDS             1272..1286
                     /codon_start=1
                     /note="CLinker"
                     /translation="ANEFM"
     CDS             1287..1790
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /note="Nluc"
                     /note="human codon-optimized"
                     /translation="LEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRIVLS
                     GENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVTPNM
                     IDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRLCE
                     RILA"
     polyA_signal    1913..2047
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2071..2087)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2095..2111
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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(2119..2149)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2164..2185
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2473..3061)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3232..4092)
                     /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"
     primer_bind     4746..4762
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5311..5690
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5691..5894
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 aagcttcccg gccacagcaa tcgatccatg acacaaccag tgacaagcag tcccccagtg
       61 aaaccgcatc cgcatccgag tccgataccg ataaagattc tgaatcggag tgagtgccgc
      121 gtccgagagc gttccctgtc cacgtccacc atcggcggag caggtgtgcc tgaggcctct
      181 agaccaccat ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc
      241 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
      301 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct
      361 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcacc aagcaccccg
      421 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga
      481 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca
      541 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc
      601 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc
      661 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact
      721 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc
      781 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct
      841 ccgaaggacg tcattctaca ggccgccata tcgatatgaa aactgtgaaa gtgaatataa
      901 caacccctga tgggccagtc tacgacgctg atatcgagat ggtgtccgtg cgggccgaga
      961 gtggtgatct cggcatcctc cccggtcaca ttcccacaaa ggccccactg aagatcggag
     1021 ctgtgcggct gaagaaggac ggccaaaccg agatggtcgc agtctcaggc ggcactgttg
     1081 aagtgcggcc tgaccacgtt accattaatg ctcaagccgc tgaaacagcc gaaggaatcg
     1141 acaaagagag agcagaagcc gcaagacaga gggcccagga gcggctgaac tctcaatccg
     1201 atgacaccga tattcgccgg gccgagctgg cactgcagag ggccctgaac agactggacg
     1261 tggctgggaa ggcaaacgaa ttcatgctgg aggactttgt tggggactgg cgacagacag
     1321 ccggctacaa cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc
     1381 tcggggtgtc cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga
     1441 tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg
     1501 aaaaaatttt taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact
     1561 atggcacact ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt
     1621 atgaaggcat cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg
     1681 gcaacaaaat tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa
     1741 ccatcaacgg agtgaccggc tggcggctgt gcgaacgcat tctggcctaa actcgacgaa
     1801 ttcattgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     1861 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     1921 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     1981 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     2041 ctggatccgt accgagctcg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2101 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2161 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2221 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2281 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2341 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2401 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2461 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2521 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2581 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2641 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     2701 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2761 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2821 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2881 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     2941 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3001 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3061 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3121 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3181 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3241 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3301 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3361 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3421 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3481 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3541 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3601 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3661 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3721 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3781 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3841 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3901 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3961 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4021 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4081 ttgaatactc atactcttcc tttttcaata agcggccgcg gccatgccgg ccactagtct
     4141 cgagttatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4201 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4261 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4321 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     4381 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     4441 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     4501 tgagagtgca ccatatgaag acgtcgcctc ctcactactt ctggaatagc tcagaggccg
     4561 aggcggcctc ggcctctgca taaataaaaa aaattagtca gcgcgccatt cgccattcag
     4621 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     4681 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     4741 acgttgtaaa acgacggcca gtgccatctg cagtgaataa taaaatgtgt gtttgtccga
     4801 aatacgcgtt tgagatttct gtcccgacta aattcatgtc gcgcgatagt ggtgtttatc
     4861 gccgatagag atggcgatat tggaaaaatc gatatttgaa aatatggcat attgaaaatg
     4921 tcgccgatgt gagtttctgt gtaactgata tcgccatttt tccaaaagtt gatttttggg
     4981 catacgcgat atctggcgat acgcttatat cgtttacggg ggatggcgat agacgccttt
     5041 ggtgacttgg gcgattctgt gtgtcgcaaa tatcgcagtt tcgatatagg tgacagacga
     5101 tatgaggcta tatcgccgat agaggcgaca tcaagctggc acatggccaa tgcatatcga
     5161 tctatacatt gaatcaatat tggccattag ccatattatt cattggttat atagcataaa
     5221 tcaatattgg ctattggcca ttgcatacgt tgtatccata tcataatatg tacatttata
     5281 ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt
     5341 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     5401 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     5461 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     5521 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     5581 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     5641 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     5701 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     5761 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     5821 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     5881 atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
     5941 ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccgggaa cggtgcattg
     6001 gaacgcggat tccccgtgcc aagagtgacg taagtaccgc ctatagagtc tataggccca
     6061 cccccttggc ttcttatgca tgctatactg tttttggctt ggggtctata cacccccgct
     6121 tcctcatgtt ataggtgatg gtatagctta gcctataggt gtgggttatt gaccattatt
     6181 gaccactccc ctattggtga cgatactttc cattactaat ccataacatg gctctttgcc
     6241 acaactctct ttattggcta tatgccaata cactgtcctt cagagactga cacggactct
     6301 gtatttttac aggatggggt ctcatttatt atttacaaat tcacatatac aacaccaccg
     6361 tccccagtgc ccgcagtttt tattaaacat aacgtgggat ctccacgcga atctcgggta
     6421 cgtgttccgg acatgggctc ttctccggta gcggcggagc ttctacatcc gagccctgct
     6481 cccatgcctc cagcgactca tggtcgctcg gcagctcctt gctcctaaca gtggaggcca
     6541 gacttaggca cagcacgatg cccaccacca ccagtgtgcc gcacaaggcc gtggcggtag
     6601 ggtatgtgtc tgaaaatgag ctcggggagc gggcttgcac cgctgacgca tttggaagac
     6661 ttaaggcagc ggcagaagaa gatgcaggca gctgagttgt tgtgttctga taagagtcag
     6721 aggtaactcc cgttgcggtg ctgttaacgg tggagggcag tgtagtctga gcagtactcg
     6781 ttgctgccgc gcgcgccacc agacataata gctgacagac taacagactg ttcctttcca
     6841 tgggtctttt ctgcagtcac cgtccttgac acg
//
