LOCUS       sequence-233784-        9076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd gen transfer vector. HLA-A0201 linked to b2-microglobulin and 
            CD28-CD3z signaling domain. Contains the Mart1 epitope 
            ""ELAGIGILTV"" cloned via BsmBI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9076)
  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 9076)
  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-A0201-Mart1-SABR
FEATURES             Location/Qualifiers
     source          1..9076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..33
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     17..33
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     52..72
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        54..72
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        173..552
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        553..756
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     707..727
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             771..951
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    998..1123
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1616..1849
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2034..2078
                     /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    2375..2492
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             5132..5197
                     /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             5282..5311
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     LTR             5694..5927
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5999..6120
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6036..6055)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6090..6109
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6160..6295
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6315..6334)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6316..6334)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6344..6361)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6344..6360)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6353..6375)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6502..6957
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6589..6608)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6799..6820
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6983..7087
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7088..7156,7157..7948)
                     /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(7306..7325)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8119..8707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8608..8627
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8861..8878
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8995..9016
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9031..9036,9037..9054,9055..9061)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(9069..9076,1..9)
                     /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     join(9074..9076,1..20)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
       61 cctcactaaa gggaacaaaa gctggagctg caagcttggc cattgcatac gttgtatcca
      121 tatcataata tgtacattta tattggctca tgtccaacat taccgccatg ttgacattga
      181 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      241 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      301 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      361 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      421 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      481 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      541 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      601 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      661 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      721 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg gggtctctct
      781 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
      841 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
      901 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
      961 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
     1021 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     1081 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     1141 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     1201 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     1261 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     1321 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     1381 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     1441 taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg
     1501 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1561 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1621 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     1681 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1741 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1801 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1861 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1921 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     1981 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     2041 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     2101 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     2161 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     2221 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     2281 gtggagagag agacagagac agatccattc gattagtgaa cggatctcga cggtatcgat
     2341 aagctaattc acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     2401 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     2461 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     2521 cagtttggga attagcttga tcgattagtc caatttgtta aagacaggat atcagtggtc
     2581 caggctctag ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag
     2641 atagaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     2701 aggtttggca agctaggatc aaggttagga acagagagac agcagaatat gggccaaaca
     2761 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagtt ggaacagcag
     2821 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     2881 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     2941 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     3001 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     3061 tcactcggcg cgatctagat ctcgaatcga attcatggcg acgggttcaa gaacttccct
     3121 acttcttgca tttggcctgc tttgtttgcc gtggttacag gagggctcgg cagagctggc
     3181 cggcatcggc atcctgaccg tgggatgcgg agggtccggc gggggaggta gcggtggtgg
     3241 aggaagcatc cagcgtactc caaagattca ggtttactca cgtcatccag cagagaatgg
     3301 aaagtcaaat ttcctgaatt gctatgtgtc tgggtttcat ccatccgaca ttgaagttga
     3361 cttactgaag aatggagaga gaattgaaaa agtggagcat tcagacttgt ctttcagcaa
     3421 ggactggtct ttctatctct tgtactacac tgaattcacc cccactgaaa aagatgagta
     3481 tgcctgccgt gtgaaccatg tgactttgtc acagcccaag atagttaagt gggatcgaga
     3541 catgggaggt ggtggcagcg gtggaggggg ctccggcggt gggggatccg gcggcggcgg
     3601 aagcggctct cactccatga ggtatttctt cacatccgtg tcccggcccg gccgtgggga
     3661 gccccgcttc atcgcagtgg gctacgtgga cgacacgcag ttcgtgcggt tcgacagcga
     3721 cgccgcgagc cagaggatgg agccgcgggc gccgtggata gagcaggagg gtccggagta
     3781 ttgggacggg gagacacgga aagtgaaggc ccactcacag actcaccgag tggacctggg
     3841 gaccctgcgc ggctgctaca accagagcga ggccggttct cacaccgtcc agaggatgta
     3901 tggctgcgac gtggggtcgg actggcgctt cctccgcggg taccaccagt acgcctacga
     3961 cggcaaggat tacatcgccc tgaaagagga cctgcgctct tggaccgcgg cggacatggc
     4021 agctcagacc accaagcaca agtgggaggc ggcccatgtg gcggagcagt tgagagccta
     4081 cctggagggc acgtgcgtgg agtggctccg cagatacctg gagaacggga aagaaacact
     4141 gcagcgcacg gacgccccca aaacgcatat gactcaccac gctgtctctg accatgaagc
     4201 caccctgagg tgctgggccc tgagcttcta ccctgcggag atcacactga cctggcagcg
     4261 ggatggggag gaccagaccc aggacacgga gctcgtggag accaggcctg caggggatgg
     4321 aaccttccag aagtgggcgg ctgtggtggt gccttctgga caggagcaga gatacacctg
     4381 ccatgtgcag catgagggtt tgcccaagcc cctcaccctg agatgggagc cgtcttccca
     4441 gcccaccatc cccatcgtgg gcatcattgc tggcctggtt ctctttggag ctgtgatcac
     4501 tggagctgtg gtcgctgctg tgatgtggag gaggaagagc tcaggtgggg aaggggtgaa
     4561 ggatagaaaa ggagggagtt acactcaggc tgcaagcagt gacagtgccc agggctctga
     4621 tgtgtctctc acagcttgta aagtgcggag caagaggtct cgcggtgggc attccgacta
     4681 catgaacatg acccctagaa ggcctggccc aaccagaaag cactaccagc catacgcccc
     4741 tcccagagat ttcgccgctt atcgaagcgt gaagttctcc cgaagcgcag atgccccagc
     4801 ctatcagcag ggacagaatc agctgtacaa cgagctgaac ctgggaagac gggaggaata
     4861 cgatgtgctg gacaaaaggc ggggcagaga tcctgagatg ggcggcaaac caagacggaa
     4921 gaacccccag gaaggtctgt ataatgagct gcagaaagac aagatggctg aggcctactc
     4981 agaaatcggg atgaagggcg aaagaaggag aggaaaaggc cacgacggac tgtaccaggg
     5041 gctgagtaca gcaacaaaag acacctatga cgctctgcac atgcaggctc tgccaccaag
     5101 aagggcaaaa cgttcgggtt cgggtgcgcc agtaaagcag acattaaact ttgatttgct
     5161 gaaacttgca ggtgatgtag agtcaaatcc aggtccaggg gcaggtgcca ccggccgcgc
     5221 catggacggg ccgcgcctgc tgctgttgct gcttctgggg gtgtcccttg gaggtgccaa
     5281 ggagcagaag ctgatcagcg aggaggacct ggagatccct ggccgttgga ttacacggtc
     5341 cacaccccca gagggctcgg acagcacagc ccccagcacc caggagcctg aggcacctcc
     5401 agaacaagac ctcatagcca gcacggtggc aggtgtggtg accacagtga tgggcagctc
     5461 ccagcccgtg gtgacccgag gcaccaccga caacctcatc cctgtctatt gctccatcct
     5521 ggctgctgtg gttgtgggtc ttgtggccta catagccttc aagaggtgag aattcgttaa
     5581 cctcgaggga tcccccgggg tcgactgatc aaattcgagc tcggtacctt taagaccaat
     5641 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     5701 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     5761 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     5821 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     5881 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     5941 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     6001 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6061 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6121 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
     6181 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6241 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg
     6301 acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     6361 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     6421 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     6481 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6541 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     6601 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     6661 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     6721 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6781 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     6841 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     6901 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     6961 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     7021 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     7081 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     7141 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     7201 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     7261 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     7321 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     7381 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7441 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7501 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7561 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7621 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7681 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7741 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7801 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7861 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7921 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7981 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     8041 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     8101 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     8161 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     8221 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     8281 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     8341 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     8401 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     8461 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     8521 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     8581 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     8641 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     8701 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     8761 tcacatcttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     8821 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     8881 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     8941 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     9001 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     9061 gtgtggaatt gtgagc
//
