LOCUS       Exported                9076 bp ds-DNA     circular SYN 13-MAY-2021
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    pCCLc-MND-A0201-Mart1-SABR
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, 
  TITLE     T cell antigen discovery via signaling and antigen-presenting 
            bifunctional receptors
  JOURNAL   Nature Methods (2019) volume 16, pages 191–198.
REFERENCE   2  (bases 1 to 9076)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature 
            Methods (2019) volume 16, pages 191–198."
FEATURES             Location/Qualifiers
     source          1..9076
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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     6222..6241
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     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             7088..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        9031..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"