LOCUS       sequence-322899-        5641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual CMV promoter plasmid to express TVA and TurboRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5641)
  AUTHORS   Faulkner RL, Wall NR, Callaway EM, Cline HT
  TITLE     Application of Recombinant Rabies Virus to Xenopus Tadpole Brain.
  JOURNAL   eNeuro. 2021 Jun 7;8(4):ENEURO.0477-20.2021. doi: 
            10.1523/ENEURO.0477-20.2021.
  PUBMED    34099488
REFERENCE   2  (bases 1 to 5641)
  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"; doi: 
            "10.1523/ENEURO.0477-20"; journalName: "eNeuro"; date: "2021-06-7- 
            7"; volume: "8"; issue: "4"; pages: "ENEURO.0477-20.2021"
COMMENT     Sequence Label: CMV::TVA/tRFP
FEATURES             Location/Qualifiers
     source          1..5641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    484..618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(710..729)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(711..729)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(736..753)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(736..752)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(745..767)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      894..1349
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(981..1000)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1191..1212
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1375..1479
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1480..1548,1549..2340)
                     /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(1698..1717)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2511..3099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3000..3019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3253..3270
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3387..3408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3423..3428,3429..3446,3447..3453)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3461..3477
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     3466..3488
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3485..3501
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3485..3501
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3522..3540
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    3644..3765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3796..4491)
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     promoter        4657..5641
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccatggcg cggctgctgc ccgcgctgct gctgctgctg
      121 ctgcccggta acgtgaccgg taacgggtcc ggtaacggtt ctttgtcccg ttgccccccc
      181 ggtcagttcc gctgctcgga gccgcccggt gcccacgggg agtgttaccc gcaggactgg
      241 ctgtgcgacg gacaccccga ctgcgacgac gggcgggacg agtggggctg cgggaccagc
      301 gcgacccccg cggtgcccac ggacaacggc acagaggctc ccactgtccc tgctcctgga
      361 cgtgctctgc cagccaggaa tcacggccgc atgtggatgc tgatcactgc agggatcttt
      421 tgctgtgagc tggtgagatg ggactgagaa ttcaaggcct ctcgagcctc tagataatac
      481 gtagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
      541 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
      601 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
      661 gaggtgtggg aggtttttta attcttgcgg ccgcttggta cccaattcgc cctatagtga
      721 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
      781 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
      841 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
      901 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
      961 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1021 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1081 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     1141 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1201 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     1261 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1321 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     1381 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1441 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1501 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1561 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1621 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1681 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1741 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1801 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1861 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1921 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1981 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2041 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2101 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2161 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2221 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2281 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2341 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2401 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2461 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2521 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2581 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2641 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2701 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2761 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2821 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2881 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2941 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3001 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3061 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3121 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3181 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3241 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3301 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3361 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     3421 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     3481 cacacaggaa acagctatga ccatgattac gccaagctcg aaattaaccc tcactaaagg
     3541 gaacaaaagc tggaggaacc ctgcaggaag ggcccaagaa ttaaaaaacc tcccacacct
     3601 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3661 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3721 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggattta
     3781 cgtattgcta gcgcatcatc tgtgccccag tttgctaggg aggtcgcagt acttggccac
     3841 agccatctcg tgctgctcga cgtaggtctc tttgtcggcc tccttgattc tttccagtct
     3901 gtggtccacg aagtggaagc cgggcatctt gaggttctta gcgggtttct tggatctgta
     3961 tgtggtcttg aaggagcagt gcaggtagcc cccgcccacg agcttcaggg ccatctggct
     4021 gtggcctctc aggccgccgt cagcggggta cagcatctcg gtgttggcct cccagccgcg
     4081 tgttttcttc tgcatcacag ggccgttgga tgggaagttc accccgttga tcttgacgtt
     4141 gtagatgatg cagccgttct ggaagctggt gtcctgggta gcggtcagca cgcccccgtc
     4201 ttcgtatgtg gtgattctct cccatgtgaa gccctcaggg aaggactgct taaagaagtc
     4261 ggggatgccc tgggtgtggt tgatgaaggc tttgctgccg tacatgaagc tggtagccag
     4321 gatgtcgaag gcgaagggga gagggccgcc ctcgaccacc ttgatcttca tggtctgggt
     4381 gccctcgtag ggcttgcctt cgccctcgga tgtgcacttg aagtggtggt tgttcacggt
     4441 gccctccatg tacagcttca tgtgcatgtt ctccttgatc agctcgctca tggtggcgag
     4501 atctgctccg acgtccccag gcagaatggc ggttccctaa acgagcattg cttatataga
     4561 cctcccatta ggcacgccta ccgcccattt acgtcaatgg aacgcccatt tgcgtcattg
     4621 cccctcccca ttgacgtcaa tggggatggg cgcgccgacc atagccaatt caatatggcg
     4681 tatatggact catgccaatt caatatggtg gatctggacc tgtgccaatt caatatggcg
     4741 tatatggact cgtgccaatt caatatggtg gatctggacc ccagccaatt caatatggcg
     4801 gacttggcac catgccaatt caatatggcg gacctggcac tgtgccaact ggggaggggt
     4861 ctacttggca cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag tccgccatat
     4921 tgaattggca tggtgccaat aatggcggcc atattggcta tatgccagga tcaatatata
     4981 ggcaatatcc aatatggccc tatgccaata tggctattgg ccaggttcaa tactatgtat
     5041 tggccctatg ccatatagta ttccatatat gggttttcct attgacgtag atagcccctc
     5101 ccaatgggcg gtcccatata ccatatatgg ggcttcctaa taccgcccat agccactccc
     5161 ccattgacgt caatggtctc tatatatggt ctttcctatt gacgtcatat gggcggtcct
     5221 attgacgtat atggcgcctc ccccattgac gtcaattacg gtaaatggcc cgcctggctc
     5281 aatgcccatt gacgtcaata ggaccaccca ccattgacgt caatgggatg gctcattgcc
     5341 cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg cccacttggc
     5401 agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga agtacgccag
     5461 ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg atggctgcca
     5521 agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg ttccattgac
     5581 gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc gtttagggaa
     5641 c
//
