LOCUS       sequence-180613-        8195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of mTagBFP2-Dectin1A-N-10 using PGK promoter
            in a lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8195)
  TITLE     Torok-Storb lab lentiviral fluorescent protein
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8195)
  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            "Unpublished"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: pRRlsinPGK_dectintagBFP2_WPRE
FEATURES             Location/Qualifiers
     source          1..8195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..46)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(200..788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(280..299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(959..1750,1751..1819))
                     /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     1582..1601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1820..1924)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1950..2405)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2087..2108)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2299..2318
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2532..2554
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2546..2563
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2547..2563
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2573..2592
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2573..2591
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2612..2747
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2787..2908
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2798..2817)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2852..2871
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             2980..3213
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3300..3888
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3406..3417
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3815..3835
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3950..4660)
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     regulatory      4657..4666
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5428..5437
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5452..5962)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5515..5533)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    6008..6125
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(6392..6436)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    6621..6854
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    7347..7472
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             7519..7699
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(7700..7926)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(7954..7974)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7954..7972)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7993..8009)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7993..8009)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8006..8028)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8017..8033
                     /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)."
     promoter        complement(join(8041..8047,8048..8065,8066..8071))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
       61 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1921 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     1981 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     2041 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     2101 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     2161 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     2221 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2281 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     2341 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2401 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     2461 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     2521 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
     2581 ctcactatag ggcgaattgg gtacgtccct aggcctccaa aaaagcctcc tcactacttc
     2641 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     2701 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     2761 ggcgggatag ctagagccag acatgataag atacattgat gagtttggac aaaccacaac
     2821 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2881 aaccattata agctgcaata aacaagttcc tctcactctc tgatattcat ttctttgcaa
     2941 gttataaata ctgaataata agatgacatg aactactact gctagagatt ttccacactg
     3001 actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta
     3061 cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag
     3121 agagctccca ggctcagatc tggtctaacc agagagaccc agtacaagca aaaagcagat
     3181 cttgtcttcg ttgggagtga attagccctt ccagtccccc cttttctttt aaaaagtggc
     3241 taagatctac agctgccttg taagtcattg gtcttaaagg taccgagctc gaattccagg
     3301 cggggaggcg gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa gacgcggaag
     3361 aggccgcaga gccggcagca ggccgcggga aggaaggtcc gctggattga gggccgaagg
     3421 gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca acacaggcga gcagccatgg
     3481 aaaggacgtc agcttccccg acaacaccac ggaattgtca gtgcccaaca gccgagcccc
     3541 tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaatag ggagggggaa
     3601 agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg ccccaaccag tgggggttgc
     3661 gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca acgggccaca actcctcata
     3721 aagagacagc aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat
     3781 agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt
     3841 aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattgt cgactgcagt
     3901 tcgaacccgg gcatatgtct agaaccggtg gatcagtcgc ggccgcttaa ttaagcttgt
     3961 gccccagttt gctagggagg tcgcagtatc tggccactgc cacctcgtgc tgctcgacgt
     4021 aggtctcgtt gttggcctcc ttgattcttt ccagtctgta gtccacatag tagacgccag
     4081 gcatcttgag gttcttagcg ggtttcttgg atctatatgt ggtcttggcg tttgcgatca
     4141 gatggctccc gcccacgagc ttcagggcca tgtcgtttct gccttccagg ccgccgtcag
     4201 cggggtacag cgtctcggtg aaggcctccc agccgagtgt tttcttctgc atcacagggc
     4261 cgttggatgt gaagttcacc cctctgatct tgacgttgta gatgaggcag ccgtcctgga
     4321 ggctggtgtc ctgggtagcg gtcagcacgc ccccgtcttc gtatgtggtg actctctccc
     4381 atgtgaagcc ctcagggaag gactgcttga agaagtcggg gatgccctgg gtgtggttga
     4441 tgaaggtctt gctgccgtag aggaagctag tagccaggat gtcgaaggcg aaggggagag
     4501 ggccgccctc gaccaccttg attctcatgg tctgggtgcc ctcgtagggc ttgccttcgc
     4561 cctcggatgt gcacttgaag tgatggttgt ccacggtgcc ctccatgtac agcttcatgt
     4621 gcatgttctc cttaatcagc tcttcgccct tagacaccat ggtggcgacc ggtggatccc
     4681 ctccggagct cattgaaaac ttcttctcac aaatactata tgagggcaca ctacacagtt
     4741 ggtcataaat gactgacacg tgaatccata cacaatttgg agatgggttt tcttgggtag
     4801 ctgtggttct gatctgaaat aagttagaag agaatgttga tccatcctcc cagagccatg
     4861 gtacttcagt ctggggccga gaaaggccta tccaaaatga attatcaggt tgggaagaca
     4921 cttgttttac tataaatccc aattcatttg agctgtctat ctttaggaga ttagagccca
     4981 gttgccagca ttgtctttta cttccatccc aggaatttag tgacatgctg aatagataac
     5041 agctcttctc atatataatc caattaggag gacaagggct ggaaagaacc cctgtggttt
     5101 tgacagcttt ggtaggagtc acactgtctt ctaaagatga ttgtgtgggt tgactgtggt
     5161 tctctttatt tcttgataga aagtagccat tctccaatgt gttgcttcct gaattggatc
     5221 tccaaatagc catggtaccc aggaccacag ctatcaccag tattaccaag cataggattc
     5281 ccaaaattac agcaatgagg cgccaaggag gagatgcagc acacgatcct ttctctgaaa
     5341 caacagctat cctggtattg ctttgagagt cgaagtgtaa ttgagtatat ccatcttcat
     5401 ccaaattttc taaatcagga tgatattcca tggtggcgct agcagatccc cctggggaga
     5461 gaggtcggtg attcggtcaa cgagggagcc gactgccgac gtgcgctccg gaggcttgca
     5521 gaatgcggaa caccgcgcgg gcaggaacag ggcccacact accgccccac accccgcctc
     5581 ccgcaccgcc ccttcccggc cgctgctctc ggcgcgccct gctgagcagc cgctattggc
     5641 cacagcccat cgcggtcggc gcgctgccat tgctccctgg cgctgtccgt ctgcgagggt
     5701 actagtgaga cgtgcggctt ccgtttgtca cgtccggcac gccgcgaacc gcaaggaacc
     5761 ttcccgactt aggggcggag caggaagcgt cgccgggggg cccacaaggg tagcggcgaa
     5821 gatccgggtg acgctgcgaa cggacgtgaa gaatgtgcga gacccagggt cggcgccgct
     5881 gcgtttcccg gaaccacgcc cagagcagcc gcgtccctgc gcaaacccag ggctgccttg
     5941 gaaaaggcgc aaccccaacc ccgtggaatt cgatatcaag cttctcgagg ctagtctcgt
     6001 gatcgataaa attttgaatt tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg
     6061 ttgctattat gtctactatt ctttcccctg cactgtaccc cccaatcccc ccttttcttt
     6121 taaaagttaa ccgataccgt cgagatccgt tcactaatcg aatggatctg tctctgtctc
     6181 tctctccacc ttcttcttct attccttcgg gcctgtcggg tcccctcggg gttgggaggt
     6241 gggtctgaaa cgataatggt gaatatccct gcctaactct attcactata gaaagtacag
     6301 caaaaactat tcttaaacct accaagcctc ctactatcat tatgaataat tttatatacc
     6361 acagccaatt tgttatgtta aaccaattcc acaaacttgc ccatttatct aattccaata
     6421 attcttgttc attcttttct tgctggtttt gcgattcttc aattaaggag tgtattaagc
     6481 ttgtgtaatt gttaatttct ctgtcccact ccatccaggt cgtgtgattc caaatctgtt
     6541 ccagagattt attactccaa ctagcattcc aaggcacagc agtggtgcaa atgagttttc
     6601 cagagcaacc ccaaatcccc aggagctgtt gatcctttag gtatctttcc acagccagga
     6661 ttcttgcctg gagctgcttg atgccccaga ctgtgagttg caacagatgc tgttgcgcct
     6721 caatagccct cagcaaattg ttctgctgct gcactatacc agacaataat tgtctggcct
     6781 gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc tgctgctccc aagaacccaa
     6841 ggaacaaagc tcctattccc actgctcttt tttctctctg caccactctt ctctttgcct
     6901 tggtgggtgc tactcctaat ggttcaattt ttactacttt atatttatat aattcacttc
     6961 tccaattgtc cctcatatct cctcctccag gtctgaagat cagcggccgc ttgctgtgcg
     7021 gtggtcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt
     7081 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt
     7141 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct
     7201 gatgtttcta acaggccagg attaactgcg aatcgttcta gctccctgct tgcccatact
     7261 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc
     7321 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct
     7381 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc
     7441 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag
     7501 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     7561 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     7621 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     7681 ggtctaacca gagagacccg tttatgtatc gagctaggca cttaaataca atatctctgc
     7741 aatgcggcaa ttcagtggtt cgtccaatcc atgtcagacc cgtctgttgc cttcctaata
     7801 aggcacgatc gtaccacctt acttccacca atcggcatgc acggtgcttt ttctctcctt
     7861 gtaaggcatg ttgctaactc atcgttacca tgttgcaaga ctacaagagt attgcataag
     7921 actacattaa gcttgcagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     7981 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     8041 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     8101 caccttaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     8161 gcattaatga atcggccaac gcgcggggag aggcg
//
