LOCUS       sequence-205455-        5449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CBR2opt in pF4Ag expression vector with T7 and CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5449)
  AUTHORS   Hall MP, Woodroofe CC, Wood MG, Que I, Van't Root M, Ridwan Y, Shi 
            C, Kirkland TA, Encell LP, Wood KV, Lowik C, Mezzanotte L
  TITLE     Click beetle luciferase mutant and near infrared naphthyl-luciferins
            for improved bioluminescence imaging.
  JOURNAL   Nat Commun. 2018 Jan 9;9(1):132. doi: 10.1038/s41467-017-02542-9.
  PUBMED    29317625
REFERENCE   2  (bases 1 to 5449)
  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: "Nat 
            Commun."; date: "2018-01-9"; volume: "9(1):132. doi"; pages: " 
            10.1038/s41467-017-02542-9"
COMMENT     Sequence Label: ATG-1929
FEATURES             Location/Qualifiers
     source          1..5449
                     /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"
     primer_bind     complement(2821..2839)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2876..2997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2887..2906)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2941..2960
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3099..3146
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3286..3390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3391..3459,3460..4251)
                     /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(3609..3628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4409..4997)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4489..4508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5113..5396
                     /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 catggttaag agagaaaaaa atgttattta tggtcctgaa cctctgcatc ctctggaaga
     1141 tctcactgca ggagagatgc tgtttcgcgc cctgcgcaag cactctcact tgccgcaggc
     1201 tctggtcgac gtggtgggtg acgagtccct gagctacaag gagttcttcg aggcgactgt
     1261 cctgctggca cagagtctcc ataactgcgg ctacaagatg aacgatgtgg tttccatctg
     1321 cgcagagaac aacactagat tctttatccc agtgatcgct gcttggtaca tcggcatgat
     1381 tgtcgcacca gtcaatgaat cttacattcc agatgaactg tgtaaggtta tgggaatttc
     1441 caaaccgcag atcgtgttca caactaagaa catcctgaac aaggttctgg aagtgcagag
     1501 tcggaccaat ttcatcaagc gtattatcat cctggacaca gtggagaaca tccacggttg
     1561 cgaatccctc cccaacttca tctctcgtta tagcgatggc aacatcgcta atttcaaacc
     1621 cctgcatttc gatcctgttg aacaggttgc cgccatcctg tgctcctctg ggaccaccgg
     1681 actgccgaaa ggcgtgatgc agacccatca gaacatctgc gtgaggctga tccacgccct
     1741 ggacccacgc tatggaactc agctgatccc aggagtcact gtgctggttt acctcccttt
     1801 ttttcacgcc ttcggtttcc acattaccct cgggtacttt atggtcggcc tccgcgttat
     1861 catgttccgt cgctttgacc aggaggcttt cctgaaggcc atccaggatt atgaggtgag
     1921 gagtgtgatt aacgtcccaa gcgtgatcct gttcctgtct aagtcccctc tggttgacaa
     1981 gtacgatctg agctccctgc gtgagctgtg ctgcggggct gcaccactgg ccaaagaagt
     2041 cgcggaagtg gctgccaaac gcctgaacct gccgggtatc tcctgtggct tcggcctgac
     2101 agagagcaca tccgctatca tccagacact gcgggatgag tttaagtccg gttccctggg
     2161 ccgggttaca cccctgatgg cagccaagat cgccgaccgc gagactggga aggccctcgg
     2221 acctaaccag gtcggagagc tgtgtattaa gggccccatg gtgtccaagg gatatgtgaa
     2281 caacgtggaa gctacaaagg aggcgattga tgatgacgga tggctccata gcggagactt
     2341 tgggtactat gacgaggacg agcacttcta tgtggtcgac aggtacaagg agctgatcaa
     2401 gtacaagggc tcccaggtgg ccccagctga actggaggag attctgttga agaacccttg
     2461 tatcagagat gtggctgttg tggggattcc tgacctcgaa gcaggtgagc tgccctccgc
     2521 tttcgtggtg aagcagcctg gtactgagat caccgccaaa gaggtgtacg attacctggc
     2581 tgagcgcgtg tcccacacca aatacctccg cggcggggtc cgtttcgtgg actccattcc
     2641 caggaacgtg accggcaaga tcacccgcaa ggagctgctg aagcagctgc tggttaaggc
     2701 tggcggcgtt taaacgaatt cgagctcggt acccggggat cctctagagt cgacctgcag
     2761 gcatgcaagc tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     2821 ccgctgagca ataactagca taaccccttg gggcggccgc ttcgagcaga catgataaga
     2881 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2941 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3001 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttta
     3061 agcaagtaaa acctctacaa atgtggtaaa atcgaattct agcataaccc cttggggcct
     3121 ctaaacgggt cttgaggggt tttttggatc ctctttgcgc ttgcgttttc ccttgtccag
     3181 atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac
     3241 gtaatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3301 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3361 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3421 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3481 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3541 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttc
     3601 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3661 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3721 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3781 actgcggcca acttacttct gacaactatc ggaggaccga aggagctaac cgcttttttg
     3841 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3901 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3961 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4021 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4081 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4141 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4201 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta attcgaaatg
     4261 accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca
     4321 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4381 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4441 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4501 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4561 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4621 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4681 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4741 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4801 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4861 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4921 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4981 gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5041 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5101 tccttttata gtccggaaat acaggaacgc acgctggatg gcccttcgct gggatggtga
     5161 aaccatgaaa aatggcagct tcagtggatt aagtgggggt aatgtggcct gtaccctctg
     5221 gttgcatagg tattcatacg gttaaaattt atcaggcgcg attgcggcag tttttcgggt
     5281 ggtttgttgc catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg
     5341 cgtacaatta agggattatg gtaaatccac ttactgtctg ccctcgtagc catcgagata
     5401 aaccgcagta ctccggccac gatgcgtccg gcgtagagga tcgagatct
//
