LOCUS       sequence-233786-        9049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd gen transfer vector. HLA-B2705 linked to b2-microglobulin and 
            CD28-CD3z signaling domain. Contains the HIV(gag) KK10 epitope 
            ""KRWIILGLNK"" cloned via BsmBI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9049)
  AUTHORS   Joglekar AV, Leonard MT, Jeppson JD, Swift M, Li G, Wong S, Peng S, 
            Zaretsky JM, Heath JR, Ribas A, Bethune MT, Baltimore D
  TITLE     T cell antigen discovery via signaling and antigen-presenting 
            bifunctional receptors.
  JOURNAL   Nat Methods. 2019 Feb;16(2):191-198. doi: 10.1038/s41592-018-0304-8.
            Epub 2019 Jan 28.
  PUBMED    30700902
REFERENCE   2  (bases 1 to 9049)
  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: 
            "10.1038/s41592-018-0304-8"; journalName: "Nat Methods"; date: 
            "2019-02"; volume: "16"; issue: "2"; pages: "191-198"
COMMENT     Sequence Label: pCCLc-MND-B2705-KK10-SABR
FEATURES             Location/Qualifiers
     source          1..9049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     134..154
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        136..154
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        255..634
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        635..838
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     789..809
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             853..1033
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1080..1205
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1698..1931
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2116..2160
                     /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    2457..2574
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             5187..5252
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             5337..5366
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     LTR             5749..5982
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6054..6175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6091..6110)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6145..6164
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6215..6350
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6370..6389)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6371..6389)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6399..6416)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6399..6415)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6408..6430)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6557..7012
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6644..6663)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6854..6875
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7038..7142
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7143..7211,7212..8003)
                     /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(7361..7380)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8174..8762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8663..8682
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8916..8933
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagcttg
      181 gccattgcat acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac
      241 attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      301 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      361 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      421 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      481 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      541 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      601 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      661 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      721 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      781 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
      841 tttagtgaac cggggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
      901 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
      961 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     1021 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
     1081 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     1141 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     1201 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     1261 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     1321 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     1381 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     1441 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     1501 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     1561 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     1621 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     1681 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     1741 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     1801 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     1861 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     1921 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     1981 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     2041 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     2101 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     2161 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     2221 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     2281 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg
     2341 cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg
     2401 aacggatctc gacggtatcg ataagctaat tcacaaatgg cagtattcat ccacaatttt
     2461 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     2521 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     2581 tattacaggg acagcagaga tccagtttgg gaattagctt gatcgattag tccaatttgt
     2641 taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc accagctgaa
     2701 gcctatagag tacgagccat agatagaata aaagatttta tttagtctcc agaaaaaggg
     2761 gggaatgaaa gaccccacct gtaggtttgg caagctagga tcaaggttag gaacagagag
     2821 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     2881 ccaagaacag ttggaacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc
     2941 tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt
     3001 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     3061 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     3121 caataaaaga gcccacaacc cctcactcgg cgcgatctag atctcgaatc gaattcatgg
     3181 cgacgggttc aagaacttcc ctacttcttg catttggcct gctttgtttg ccgtggttac
     3241 aggagggctc ggcaaagagg tggatcatcc tgggcctgaa caagggatgc ggagggtccg
     3301 gcgggggagg tagcggtggt ggaggaagca tccagcgtac tccaaagatt caggtttact
     3361 cacgtcatcc agcagagaat ggaaagtcaa atttcctgaa ttgctatgtg tctgggtttc
     3421 atccatccga cattgaagtt gacttactga agaatggaga gagaattgaa aaagtggagc
     3481 attcagactt gtctttcagc aaggactggt ctttctatct cttgtactac actgaattca
     3541 cccccactga aaaagatgag tatgcctgcc gtgtgaacca tgtgactttg tcacagccca
     3601 agatagttaa gtgggatcga gacatgggag gtggtggcag cggtggaggg ggctccggcg
     3661 gtgggggatc cggcggcggc ggaagcggct cccactccat gaggtatttc cacacctccg
     3721 tgtcccggcc cggccgcggg gagccccgct tcatcaccgt gggctacgtg gacgacacgc
     3781 tgttcgtgag gttcgacagc gacgccgcga gtccgagaga ggagccgcgg gcgccgtgga
     3841 tagagcagga ggggccggag tattgggacc gggagacaca gatctgcaag gccaaggcac
     3901 agactgaccg agaggacctg cggaccctgc tccgctgcta caaccagagc gaggccgggt
     3961 ctcacaccct ccagaatatg tatggctgcg acgtggggcc ggacgggcgc ctcctccgcg
     4021 ggtaccacca ggacgcctac gacggcaagg attacatcgc cctgaacgag gacctgagct
     4081 cctggaccgc cgcggacacg gcggctcaga tcacccagcg caagtgggag gcggcccgtg
     4141 tggcggagca gctgagagcc tacctggagg gcgagtgcgt ggagtggctc cgcagatacc
     4201 tggagaacgg gaaagaaaca ctgcagcgcg cggacccccc aaagacacac gtgacccacc
     4261 accccatctc tgaccatgag gccaccctga ggtgctgggc cctgggcttc taccctgcgg
     4321 agatcacact gacctggcag cgggatggcg aggaccaaac tcaggacact gagcttgtgg
     4381 agaccagacc agcaggagat agaaccttcc agaagtgggc agctgtggtg gtgccttctg
     4441 gagaagagca gagatacaca tgccatgtac agcatgaggg gctgccgaag cccctcaccc
     4501 tgagatggga gccgtcttcc cagtccaccg tccccatcgt gggcattgtt gctggcctgg
     4561 ctgtcctagc agttgtggtc atcggagctg tggtcgctgc tgtgatgtgt aggaggaaga
     4621 gctcaggtgg aaaaggaggg agctactctc aggctgcgtg cagcgacagt gcccagggct
     4681 ctgatgtgtc tctcacagct cggagcaaga ggtctcgcgg tgggcattcc gactacatga
     4741 acatgacccc tagaaggcct ggcccaacca gaaagcacta ccagccatac gcccctccca
     4801 gagatttcgc cgcttatcga agcgtgaagt tctcccgaag cgcagatgcc ccagcctatc
     4861 agcagggaca gaatcagctg tacaacgagc tgaacctggg aagacgggag gaatacgatg
     4921 tgctggacaa aaggcggggc agagatcctg agatgggcgg caaaccaaga cggaagaacc
     4981 cccaggaagg tctgtataat gagctgcaga aagacaagat ggctgaggcc tactcagaaa
     5041 tcgggatgaa gggcgaaaga aggagaggaa aaggccacga cggactgtac caggggctga
     5101 gtacagcaac aaaagacacc tatgacgctc tgcacatgca ggctctgcca ccaagaaggg
     5161 caaaacgttc gggttcgggt gcgccagtaa agcagacatt aaactttgat ttgctgaaac
     5221 ttgcaggtga tgtagagtca aatccaggtc caggggcagg tgccaccggc cgcgccatgg
     5281 acgggccgcg cctgctgctg ttgctgcttc tgggggtgtc ccttggaggt gccaaggagc
     5341 agaagctgat cagcgaggag gacctggaga tccctggccg ttggattaca cggtccacac
     5401 ccccagaggg ctcggacagc acagccccca gcacccagga gcctgaggca cctccagaac
     5461 aagacctcat agccagcacg gtggcaggtg tggtgaccac agtgatgggc agctcccagc
     5521 ccgtggtgac ccgaggcacc accgacaacc tcatccctgt ctattgctcc atcctggctg
     5581 ctgtggttgt gggtcttgtg gcctacatag ccttcaagag gtgagaattc gttaacctcg
     5641 agggatcccc cggggtcgac tgatcaaatt cgagctcggt acctttaaga ccaatgactt
     5701 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     5761 ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     5821 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     5881 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     5941 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     6001 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     6061 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6121 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6181 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6241 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6301 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
     6361 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     6421 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     6481 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     6541 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     6601 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     6661 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     6721 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     6781 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6841 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     6901 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     6961 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     7021 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     7081 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     7141 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     7201 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     7261 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     7321 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     7381 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     7441 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     7501 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     7561 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     7621 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     7681 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     7741 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     7801 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     7861 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7921 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7981 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     8041 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     8101 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     8161 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     8221 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     8281 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     8341 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     8401 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     8461 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     8521 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     8581 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     8641 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8701 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     8761 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     8821 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     8881 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     8941 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     9001 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaat
//
