LOCUS       Exported                6071 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses hCD19.
ACCESSION   .
VERSION     .
KEYWORDS    MSCV-hCD19
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6071)
  AUTHORS   Chen J, Lopez-Moyado IF, Seo H, Lio CJ, Hempleman LJ, Sekiya T, 
            Yoshimura A, Scott-Browne JP, Rao A
  TITLE     NR4A transcription factors limit CAR T cell function in solid 
            tumours.
  JOURNAL   Nature. 2019 Mar;567(7749):530-534. doi: 10.1038/s41586-019-0985-x. 
            Epub 2019 Feb 27.
  PUBMED    30814732
REFERENCE   2  (bases 1 to 6071)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6071
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     RBS             1441..1452
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     LTR             2464..2978
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3140..3156
                     /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(3164..3194)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3209..3230
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3347..3364)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3518..4106)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3598..3617)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4277..5137)
                     /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     4900..4919
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5138..5242)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5310..5328
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5366..5388)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5488..5507
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5701..5723
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5851..5870
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6070..515
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gccaccatgc ctcccccacg actgttgttt
     1441 tttctcctct ttctgacacc aatggaggtg cgcccagaag agcctctggt ggttaaggta
     1501 gaagaagggg ataatgccgt actccaatgc cttaagggga catcagatgg accaactcaa
     1561 cagttgacct ggagcaggga gtcccctctc aaaccttttc ttaagttgtc acttggcctt
     1621 ccagggctcg gtatccatat gagacccctc gcaatctggt tgttcatctt taacgtctct
     1681 cagcagatgg gagggtttta cctttgccag ccaggaccac ctagcgaaaa ggcttggcaa
     1741 ccaggttgga ctgtcaatgt tgagggaagc ggtgagttgt ttcgctggaa cgtaagcgac
     1801 ctgggggggc tcggatgtgg attgaaaaat cgatcctccg aagggccttc ttccccttca
     1861 ggcaagttga tgtcccccaa actgtatgta tgggccaagg atcggcccga gatttgggaa
     1921 ggagaacccc catgcttgcc tccaagagac agtcttaatc agtccctttc acaggacttg
     1981 actatggccc caggtagtac cttgtggctg tcatgcggtg tgccccctga tagtgtgtct
     2041 cggggtcctc tgagctggac ccacgtccat cccaaaggac caaaatccct tctttcactc
     2101 gaattgaaag acgaccgccc tgcacgcgac atgtgggtga tggagactgg ccttttgctc
     2161 ccccgagcca cagctcagga tgcaggaaag tactactgtc atcgaggtaa cctgactatg
     2221 tcctttcacc tggaaatcac tgcacgccca gtgctttggc attggttgct cagaacagga
     2281 ggctggaagg ttagcgcagt cacattggct tatctgatct tctgtctttg tagccttgtc
     2341 ggaatactgc acctgcagcg agcccttgtg ctcagacgca agcggaagag aatgactgat
     2401 ccaacaagaa ggttctgata aataaaataa aagattttat ttagtctcca gaaaaagggg
     2461 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     2521 gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga acagagagac
     2581 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     2641 aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat
     2701 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     2761 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     2821 acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc
     2881 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     2941 tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt cttgaagttg
     3001 gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca attagccact
     3061 gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag cgcagaaaga
     3121 gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3181 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3241 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3301 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3361 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3421 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3481 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3541 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3601 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3661 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3721 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3781 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3841 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3901 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3961 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4021 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4081 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4141 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4201 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4261 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4321 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4381 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4441 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4501 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4561 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4621 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4681 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4741 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4801 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4861 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4921 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4981 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5041 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5101 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5161 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5221 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     5281 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     5341 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     5401 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     5461 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5521 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     5581 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     5641 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     5701 cccagtcacg acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg agatggcgcc
     5761 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     5821 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     5881 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagaggc gattagtcca
     5941 atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac aatatcacca
     6001 gctgaagcct atagagtacg agccatagat aaaataaaag attttattta gtctccagaa
     6061 aaagggggga a
//
