LOCUS       sequence-335154-        5470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CBG2 (Click Beetle Green 2) in E. coli and mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5470)
  AUTHORS   Zambito G, Hall MP, Wood MG, Gaspar N, Ridwan Y, Stellari FF, Shi C,
            Kirkland TA, Encell LP, Lowik C, Mezzanotte L
  TITLE     Red-shifted click beetle luciferase mutant expands the multicolor 
            bioluminescent palette for deep tissue imaging.
  JOURNAL   iScience. 2020 Dec 26;24(1):101986. doi: 10.1016/j.isci.2020.101986.
            eCollection 2021 Jan 22.
  PUBMED    33490896
REFERENCE   2  (bases 1 to 5470)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "iScience.";
            date: "2020-12-26"; volume: "24(1):101986. doi"; pages: " 
            10.1016/j.isci.2020.101986. eCollection 2021 Jan 22"
COMMENT     Sequence Label: ATG-3925 (CBG2)
FEATURES             Location/Qualifiers
     source          1..5470
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          720..852
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     835..854
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     896..915
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        896..914
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             951..968
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2705..2723)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2760..2881
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2771..2790)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2825..2844
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2983..3030
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3170..3274
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3275..3343,3344..4135)
                     /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"
     primer_bind     complement(3493..3512)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4293..4881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4373..4392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4997..5280
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatcactag aagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg
      661 cttctgacac aacagtctcg aacttaagct gcagaagttg gtcgtgaggc actgggcagg
      721 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
      781 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
      841 ttctctccac aggtgtccac tcccagttca attacagctc ttaaggctag agtattaata
      901 cgactcacta tagggaattt taactttact aaggagcgat cgccatgaaa catcaccatc
      961 accatcatgt taagagagaa aaaaatgtta tttatggtcc tgaacctctg catcctctgg
     1021 aagatctcac tgcaggagag atgctgtttc gcgccctgcg caagcactct cacttgccgc
     1081 aggctctggt cgacgtggtg ggtgacgagt ccctgagcta caaggagttc ttcgaggcga
     1141 ctgtcctgct ggcacagagt ctccataact gcggctacaa gatgaacgat gtggtttcca
     1201 tctgcgcaga gaacaacact agattcttta tcccagtgat cgctgcttgg tacatcggca
     1261 tgattgtcgc accagtcaat gaatcttaca ttccagatga actgtgtaag gttatgggaa
     1321 tttccaaacc gcagatcgtg ttcacaacta agaacatcct gaacaaggtt ctggaagtgc
     1381 agagtcggac caatttcatc aagcgtatta tcatcctgga cacagtggag aacatccacg
     1441 gttgcgaatc cctccccaac ttcatctctc gttatagcga tggcaacatc gctaatttca
     1501 aacccctgca tttcgatcct gttgaacagg ttgccgccat cctgtgctcc tctgggacca
     1561 ccggactgcc gaaaggcgtg atgcagaccc atcagaacat ctgcgtgagg ctgatccacg
     1621 ccctggaccc acgctatgga actcagctga tcccaggagt cactgtgctg gtttacctcc
     1681 ctttttttca cgccttcggt ttccacatta ccctcgggta ctttatggtc ggcctccgcg
     1741 ttatcatgtt ccgtcgcttt gaccaggagg ctttcctgaa ggccatccag gattatgagg
     1801 tgaggagtgt gattaacgtc ccaagcgtga tcctgttcct gtctaagtcc cctctggttg
     1861 acaagtacga tctgagctcc ctgcgtgagc tgtgctgcgg ggctgcacca ctggccaaag
     1921 aagtcgcgga agtggctgcc aaacgcctga acctgccggg tatccgctgt ggcttcggcc
     1981 tgacagagag cacatccgct atcatccata cactggggga tgagtttaag tccggttccc
     2041 tgggccgggt tacacccctg atggcagcca agatcgccga ccgcgagact gggaaggccc
     2101 tcggacctaa ccaggtcgga gagctgtgta ttaagggccc catggtgtcc aagggatatg
     2161 tgaacaacgt ggaagctaca aaggaggcga ttgatgatga cggatggctc catagcggag
     2221 actttgggta ctatgacgag gacgagcact tctatgtggt cgacaggtac aaggagctga
     2281 tcaagtacaa gggctcccag gtggccccag ctgaactgga ggagattctg ttgaagaacc
     2341 cttgtatcag agatgtggct gttgtgggga ttcctgacct cgaagcaggt gagctgccct
     2401 ccgctttcgt ggtgaagcag cctggtactg agatcaccgc caaagaggtg tacgattacc
     2461 tggctgagcg cgtgtcccac accaaatacc tccgcggcgg ggtccgtttc gtggactcca
     2521 ttcccaggaa cgtgaccggc aagatcaccc gcaaggagct gctgaagcag ctgctggtta
     2581 aggctggcgg cgtttaaacg aattcgagct cggtacccgg ggatcctcta gagtcgacct
     2641 gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     2701 gccaccgctg agcaataact agcataaccc cttggggcgg ccgcttcgag cagacatgat
     2761 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2821 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2881 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     2941 tttaagcaag taaaacctct acaaatgtgg taaaatcgaa ttctagcata accccttggg
     3001 gcctctaaac gggtcttgag gggttttttg gatcctcttt gcgcttgcgt tttcccttgt
     3061 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     3121 ctacgtaatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3181 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3241 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3301 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3361 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3421 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3481 tttcaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3541 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3601 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3661 taacactgcg gccaacttac ttctgacaac tatcggagga ccgaaggagc taaccgcttt
     3721 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3781 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3841 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3901 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3961 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4021 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4081 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaattcga
     4141 aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt aatacggtta
     4201 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4261 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4321 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4381 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4441 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4501 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4561 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4621 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4681 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     4741 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4801 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4861 cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4921 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4981 tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt cgctgggatg
     5041 gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc
     5101 tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg gcagtttttc
     5161 gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc
     5221 ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg tagccatcga
     5281 gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga tcttcaatat
     5341 tggccattag ccatattatt cattggttat atagcataaa tcaatattgg ctattggcca
     5401 ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga
     5461 ccgccatgtt
//
