LOCUS       sequence-345061-        8242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRIM46 tagged with iRFP702.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8242)
  TITLE     Fluorescent plasmids for live cell imaging in primary neurons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8242)
  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: 
            "Unpublished"
COMMENT     Sequence Label: iRFP-TRIM46
FEATURES             Location/Qualifiers
     source          1..8242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(11..17,18..35,36..41))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(193..210)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(364..952)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(444..463)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1123..1914,1915..1983))
                     /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     1746..1765
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     2171..2189
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2227..2249)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2349..2368
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2600..2622
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2614..2631
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2615..2631
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3197..3576
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3577..3780
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3731..3751
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3777..3801
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4682..4691
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4688..5620
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 702 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP702
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGKVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPIHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     polyA_signal    8044..8178
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8081..8100)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8135..8154
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8205..8221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8205..8221)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8218..8240)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    join(8229..8242,1..3)
                     /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)."
ORIGIN
        1 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
       61 tgagctaact acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
      121 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
      181 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      241 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      301 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      361 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      421 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      481 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      541 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      601 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      661 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      721 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      781 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      841 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      901 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      961 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1021 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1081 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1141 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1201 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1261 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1321 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1381 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1441 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1501 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1561 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1621 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1681 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1741 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1801 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1861 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1921 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1981 catactcttc ctttttcaat aagcggcctc gagttattat tgaagcattt atcagggtta
     2041 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2101 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     2161 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     2221 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     2281 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     2341 taactatgcg gcatcagagc agattgtact gagagtgcac catatgaaga cgtcgcctcc
     2401 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     2461 aattagtcag ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     2521 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     2581 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgcccgtgtc
     2641 aaggacggtg actgcagtga ataataaaat gtgtgtttgt ccgaaatacg cgttttgaga
     2701 tttctgtcgc cgactaaatt catgtcgcgc gatagtggtg tttatcgccg atagagatgg
     2761 cgatattgga aaaatcgata tttgaaaata tggcatattg aaaatgtcgc cgatgtgagt
     2821 ttctgtgtaa ctgatatcgc catttttcca aaagtgattt ttgggcatac gcgatatctg
     2881 gcgatagcgc ttatatcgtt tacgggggat ggcgatagac gactttggtg acttgggcga
     2941 ttctgtgtgt cgcaaatatc gcagtttcga tataggtgac agacgatatg aggctatatc
     3001 gccgatagag gcgacatcaa gctggcacat ggccaatgca tatcgatcta tacattgaat
     3061 caatattggc cattagccat attattcatt ggttatatag cataaatcaa tattggctat
     3121 tggccattgc atacgttgta tccatatcat aatatgtaca tttatattgg ctcatgtcca
     3181 acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     3241 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     3301 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     3361 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     3421 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     3481 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     3541 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     3601 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     3661 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     3721 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     3781 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
     3841 agacaccggg accgatccag cctccgcggc cgggaacggt gcattggaac gcggattccc
     3901 cgtgccaaga gtgacgtaag taccgcctat agagtctata ggcccacccc cttggcttct
     3961 tatgcatgct atactgtttt tggcttgggg tctatacacc cccgcttcct catgttatag
     4021 gtgatggtat agcttagcct ataggtgtgg gttattgacc attattgacc actcccctat
     4081 tggtgacgat actttccatt actaatccat aacatggctc tttgccacaa ctctctttat
     4141 tggctatatg ccaatacact gtccttcaga gactgacacg gactctgtat ttttacagga
     4201 tggggtctca tttattattt acaaattcac atatacaaca ccaccgtccc cagtgcccgc
     4261 agtttttatt aaacataacg tgggatctcc acgcgaatct cgggtacgtg ttccggacat
     4321 gggctcttct ccggtagcgg cggagcttct acatccgagc cctgctccca tgcctccagc
     4381 gactcatggt cgctcggcag ctccttgctc ctaacagtgg aggccagact taggcacagc
     4441 acgatgccca ccaccaccag tgtgccgcac aaggccgtgg cggtagggta tgtgtctgaa
     4501 aatgagctcg gggagcgggc ttgcaccgct gacgcatttg gaagacttaa ggcagcggca
     4561 gaagaagatg caggcagctg agttgttgtg ttctgataag agtcagaggt aactcccgtt
     4621 gcggtgctgt taacggtgga gggcagtgta gtctgagcag tactcgttgc tgccgcgcgc
     4681 ggccaccatg gcgcgtaagg tcgatctcac ctcctgcgat cgcgagccga tccacatccc
     4741 cggcagcatt cagccgtgcg gctgcctgct agcctgcgac gcgcaggcgg tgcggatcac
     4801 gcgcattacg gaaaatgccg gcgcgttctt tggacgcgaa actccgcggg tcggtgagct
     4861 actcgccgat tacttcggcg agaccgaagc ccatgcgctg cgcaacgcac tggcgcagtc
     4921 ctccgatcca aagcgaccgg cgctgatctt cggttggcgc gacggcctga ccggccgcac
     4981 cttcgacatc tcgctgcatc gccatgacgg tacatcgatc atcgagttcg agcctgcggc
     5041 ggccgaacag gccgacaatc cgctgcggct gacgcggcag atcatcgcgc gcaccaaaga
     5101 actgaagtcg ctcgaagaga tggccgcacg ggtgccgcgc tatctgcagg cgatgctcgg
     5161 ctatcaccgc gtgatgttgt accgcttcgc ggacgacggc tccggcaaag tgatcggcga
     5221 ggcgaagcgc agcgacctcg agagctttct cggtcagcac tttccggcgt cgctggtccc
     5281 gcagcaggcg cggctactgt acttgaagaa cgcgatccgc gtggtctcgg attcgcgcgg
     5341 catcagcagc cggatcgtgc ccgagcacga cgcctccggc gccgcgcttg atctgtcgtt
     5401 cgcgcacctg cgcagcatct cgcctatcca tctcgaattt ctgcggaaca tgggcgtcag
     5461 cgcctcgatg tcgctgtcga tcatcattga cggcacgcta tggggattga tcatctgtca
     5521 tcattacgag ccgcgtgccg tgccgatggc gcagcgcgtc gcggccgaaa tgttcgccga
     5581 cttcttatcg ctgcacttca ccgccgccca ccaccaacgc gaattcatgg ctgagggtga
     5641 ggatatgcag accttcactt ccatcatgga tgcactagtc cgcatcagta caagcatgaa
     5701 gaacatggag aaggaactgc tgtgcccagt gtgccaagag atgtacaagc agccactagt
     5761 gctgccttgt acccacaacg tgtgccaggc ctgtgcccgg gaggttttgg gccagcaggg
     5821 gtacataggc catggtgggg atccgagttc ggagcccact tctcctgctt ccaccccttc
     5881 tacccgcagc ccccgcctct ctcggagaac tctccccaag ccagatcgct tggaccggct
     5941 ccttaagtca ggctttggga cataccctgg gcggaagcgt ggtgccttgc acccccagac
     6001 gatcctattc ccgtgcccag cctgccaagg cgatgtggag ctgggggaga ggggcctttc
     6061 cgggcttttc cgaaacctga ccctagagcg agtggtggag aggtaccgcc agagtgtgag
     6121 tgtgggagga gccatcctgt gccagttgtg caagccccca ccactagagg ccaccaaggg
     6181 ctgcacagag tgtcgagcta ccttctgtaa cgaatgcttc aagctcttcc acccctgggg
     6241 cactcagaag gcgcagcatg agcccacgct acccactctc tcattccgcc ccaagggtct
     6301 gatgtgtcca gaccacaagg aagaggtgac ccactactgt aagacatgtc aacgactggt
     6361 gtgccaactc tgcagagtac ggcgtaccca cagtgggcac aagatcacac cagtgctcag
     6421 tgcctaccag gccctcaagg ataagctaac aaagagcctg gcatacatct tgggaaatca
     6481 ggacactgtg cagacccaga tttgtgagct ggaagagacc atcaggcaca ctgaggtgag
     6541 tggtcagcaa gcaaaagagg aggtgtccca gctggttcgg ggactagggg ctgtgctgga
     6601 agagaagcgg gcctcactgc ttcaggccat tgaagaatgc cagcaagagc gcttatcccg
     6661 gctcagcgcc cagatccatg aacaccagag cctgctggat ggctcgggtc tggtgggtta
     6721 tgcgcaggaa gtccttaagg aaacagacca gccttgtttt gtacaagcag ccaaacagct
     6781 gcataacagg attgcccgag ccactgaggc cctccagaca ttccggccag ctgccagctc
     6841 ctccttccgc cactgccagc tggatgtggg gcgtgagatg aagctgttga ccgagcttag
     6901 cttcctgaga gtgcccgagg cgcccgtcat agacacccaa cgcacctttg cctatgatca
     6961 aatcttcctg tgctggcggc tgccccctca ctcaccacct gcctggcatt acaccgttga
     7021 gttccggcgc acagatgtcc ccgcccagcc aggccctaca cgctggcagc gccgggaaga
     7081 ggtgaggggc accagcgccc ttcttgagaa ccccgacacg ggctctgtgt acgtgctgcg
     7141 tgtccgtggc tgcaacaagg ctggctacgg agagtacagt gaagatgtgc acctgcacac
     7201 acccccagcc cctgtcctgc acttcttcct tgatggccgc tggggtgcga gccgagagcg
     7261 gctagccatc agcaaggacc agcgagcggt gcggagtatc ccagggcttc ccttgctgct
     7321 ggctgctgag cggctgctca cgggctgcca cctgagtgtg gacgtggtct tgggtgatgt
     7381 ggctgtgacc cagggccgca gctactgggc ctgtgcggta gaccctgcct cctacttggt
     7441 caaggtgggc gtcgggctgg agagcaaact tcaggaaagc ttccagggtg ccccggatgt
     7501 gatcagcccc agatacgacc cggacagtgg gcacgacagc ggtgccgagg acgctgccgt
     7561 ggaggcgtta ccaccctttg ctttcctgac catcggcatg ggcaagatcc tgctgggttc
     7621 cggagcgagt tccaatgcgg ggctgacagg gagggatggc cccaccacca gctgcacagt
     7681 gcccctgcca ccccgcctag gtatctgttt ggactacgag cgggggcggg tctccttcct
     7741 ggatgccgtg tccttccgtg ggctgctcga gtgccccctg gactgctcag ggcctgtgtg
     7801 ccctgccttc tgcttcattg ggggtggcgc tgttcagctg caggagcccg tgggcaccaa
     7861 gcctgagagg aaagtcacca ttgggggctt tgccaaactg gactgagtcg actgcggccg
     7921 cctctagata attcattgat cataatcagc cataccacat ttgtagaggt tttacttgct
     7981 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     8041 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     8101 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     8161 tcttatcatg tctggatcga tccgtaccga gctcgcgtaa tcatgtcata gctgtttcct
     8221 gtgtgaaatt gttatccgct ca
//
