LOCUS       sequence-325762-        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        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1088..1105
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2842..2860)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2897..3018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2908..2927)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2962..2981
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3120..3167
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3307..3411
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3412..3480,3481..4272)
                     /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(3630..3649)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4430..5018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4510..4529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5134..5417
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggaattttaa ctttactaag gagcgatcgc
     1081 catgaaacat caccatcacc atcatgttaa gagagaaaaa aatgttattt atggtcctga
     1141 acctctgcat cctctggaag atctcactgc aggagagatg ctgtttcgcg ccctgcgcaa
     1201 gcactctcac ttgccgcagg ctctggtcga cgtggtgggt gacgagtccc tgagctacaa
     1261 ggagttcttc gaggcgactg tcctgctggc acagagtctc cataactgcg gctacaagat
     1321 gaacgatgtg gtttccatct gcgcagagaa caacactaga ttctttatcc cagtgatcgc
     1381 tgcttggtac atcggcatga ttgtcgcacc agtcaatgaa tcttacattc cagatgaact
     1441 gtgtaaggtt atgggaattt ccaaaccgca gatcgtgttc acaactaaga acatcctgaa
     1501 caaggttctg gaagtgcaga gtcggaccaa tttcatcaag cgtattatca tcctggacac
     1561 agtggagaac atccacggtt gcgaatccct ccccaacttc atctctcgtt atagcgatgg
     1621 caacatcgct aatttcaaac ccctgcattt cgatcctgtt gaacaggttg ccgccatcct
     1681 gtgctcctct gggaccaccg gactgccgaa aggcgtgatg cagacccatc agaacatctg
     1741 cgtgaggctg atccacgccc tggacccacg ctatggaact cagctgatcc caggagtcac
     1801 tgtgctggtt tacctccctt tttttcacgc cttcggtttc cacattaccc tcgggtactt
     1861 tatggtcggc ctccgcgtta tcatgttccg tcgctttgac caggaggctt tcctgaaggc
     1921 catccaggat tatgaggtga ggagtgtgat taacgtccca agcgtgatcc tgttcctgtc
     1981 taagtcccct ctggttgaca agtacgatct gagctccctg cgtgagctgt gctgcggggc
     2041 tgcaccactg gccaaagaag tcgcggaagt ggctgccaaa cgcctgaacc tgccgggtat
     2101 ccgctgtggc ttcggcctga cagagagcac atccgctatc atccatacac tgggggatga
     2161 gtttaagtcc ggttccctgg gccgggttac acccctgatg gcagccaaga tcgccgaccg
     2221 cgagactggg aaggccctcg gacctaacca ggtcggagag ctgtgtatta agggccccat
     2281 ggtgtccaag ggatatgtga acaacgtgga agctacaaag gaggcgattg atgatgacgg
     2341 atggctccat agcggagact ttgggtacta tgacgaggac gagcacttct atgtggtcga
     2401 caggtacaag gagctgatca agtacaaggg ctcccaggtg gccccagctg aactggagga
     2461 gattctgttg aagaaccctt gtatcagaga tgtggctgtt gtggggattc ctgacctcga
     2521 agcaggtgag ctgccctccg ctttcgtggt gaagcagcct ggtactgaga tcaccgccaa
     2581 agaggtgtac gattacctgg ctgagcgcgt gtcccacacc aaatacctcc gcggcggggt
     2641 ccgtttcgtg gactccattc ccaggaacgt gaccggcaag atcacccgca aggagctgct
     2701 gaagcagctg ctggttaagg ctggcggcgt ttaaacgaat tcgagctcgg tacccgggga
     2761 tcctctagag tcgacctgca ggcatgcaag ctgatccggc tgctaacaaa gcccgaaagg
     2821 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcggccg
     2881 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     2941 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3001 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3061 gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa aatcgaattc
     3121 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttggat cctctttgcg
     3181 cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg tcagcaccgt
     3241 ttctgcggac tggctttcta cgtaatggtt tcttagacgt caggtggcac ttttcgggga
     3301 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3361 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3421 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3481 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3541 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3601 tttccaatga tgagcacttt caaagttctg ctatgtggcg cggtattatc ccgtattgac
     3661 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3721 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3781 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaactat cggaggaccg
     3841 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3901 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3961 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4021 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4081 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4141 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4201 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4261 aagcattggt aattcgaaat gaccgaccaa gcgacgccca accggtatca gctcactcaa
     4321 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4381 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4441 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4501 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4561 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4621 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4681 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4741 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4801 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4861 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4921 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4981 gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg atcttttcta
     5041 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5101 caaaaaggat cttcacctag atccttttat agtccggaaa tacaggaacg cacgctggat
     5161 ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc ttcagtggat taagtggggg
     5221 taatgtggcc tgtaccctct ggttgcatag gtattcatac ggttaaaatt tatcaggcgc
     5281 gattgcggca gtttttcggg tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc
     5341 gctgaacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca cttactgtct
     5401 gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg
     5461 atcgagatct
//
