LOCUS       sequence-412341-        8191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid can be used to generate retrovirus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8191)
  AUTHORS   Blaeschke F, Chen YY, Apathy R, Daniel B, Chen AY, Chen PA, Sandor 
            K, Zhang W, Li Z, Mowery CT, Yamamoto TN, Nyberg WA, To A, Yu R, 
            Bueno R, Kim MC, Schmidt R, Goodman DB, Feuchtinger T, Eyquem J, 
            Jimmie Ye C, Carnevale J, Satpathy AT, Shifrut E, Roth TL, Marson A
  TITLE     Modular pooled discovery of synthetic knockin sequences to program 
            durable cell therapies.
  JOURNAL   Cell. 2023 Sep 14;186(19):4216-4234.e33. doi: 
            10.1016/j.cell.2023.08.013.
  PUBMED    37714135
REFERENCE   2  (bases 1 to 8191)
  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: 
            "10.1016/j.cell.2023.08"; journalName: "Cell"; date: "2023-09-14- 
            14"; volume: "186"; issue: "19"; pages: "4216-4234.e33"
COMMENT     Sequence Label: pIG-828_HA-GD2-28z_CAR_BATF_Retroviral
FEATURES             Location/Qualifiers
     source          1..8191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1204..1222
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1260..1282)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1382..1401
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1595..1617
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1745..1764
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             1964..2480
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     3320..3342
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3358..3374
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     3712..3728
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             4225..4278
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     LTR             6539..7053
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7225..7241
                     /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(7249..7255,7256..7273,7274..7279))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7294..7315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7432..7449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7603..8191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7683..7702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     1201 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     1261 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     1321 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     1381 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     1441 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     1501 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     1561 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     1621 gcgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     1681 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     1741 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     1801 ccacgatgcg tccggcgtag aggcgattag tccaatttgt taaagacagg atatcagtgg
     1861 tccaggctct agttttgact caacaatatc accagctgaa gcctatagag tacgagccat
     1921 agataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     1981 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact
     2041 gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac
     2101 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     2161 atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     2221 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     2281 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc
     2341 agtcctccga tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt
     2401 tgcatccgaa tcgtggactc gctgatcctt gggagggtct cctcagattg attgactgcc
     2461 cacctcgggg gtctttcatt tggaggttcc accgagattt ggagacccct gcccagggac
     2521 caccgacccc cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct
     2581 ttgtgcgtgt ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact
     2641 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt cggaacaccc ggccgcaacc
     2701 ctgggagacg tcccagggac ttcgggggcc gtttttgtgg cccgacctga gtcctaaaat
     2761 cccgatcgtt taggactctt tggtgcaccc cccttagagg agggatatgt ggttctggta
     2821 ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttgggac
     2881 cgaagccgcg ccgcgcgtct tgtctgctgc agcatcgttc tgtgttgtct ctgtctgact
     2941 gtgtttctgt atttgtctga aaatatgggc ccgggctagc ctgttaccac tcccttaagt
     3001 ttgaccttag gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc
     3061 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     3121 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     3181 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     3241 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct
     3301 ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt
     3361 tatccagccc tcactccttc tctaggcgcc cccatatggc catatgagat cttatatggg
     3421 gcacccccgc cccttgtaaa cttccctgac cctgacatga caagagttac taacagcccc
     3481 tctctccaag ctcacttaca ggctctctac ttagtccagc acgaagtctg gagacctctg
     3541 gcggcagcct accaagaaca actggaccga ccggtggtac ctcaccctta ccgagtcggc
     3601 gacacagtgt gggtccgccg acaccagact aagaacctag aacctcgctg gaaaggacct
     3661 tacacagtcc tgctgaccac ccccaccgcc ctcaaagtag acggcatcgc agcttggata
     3721 cacgccgccc acgtgaaggc tgccgacccc gggggtggac catcctctag ccctcgagcc
     3781 gccaccatgc ctcacagctc cgacagcagt gactccagct tcagccgctc tcctccccct
     3841 ggcaaacagg actcatctga tgatgtgaga agagttcaga ggagggagaa aaatcgtatt
     3901 gccgcccaga agagccgaca gaggcagaca cagaaggccg acaccctgca cctggagagc
     3961 gaagacctgg agaaacagaa cgcggctcta cgcaaggaga tcaagcagct cacagaggaa
     4021 ctgaagtact tcacgtcggt gctgaacagc cacgagcccc tgtgctcggt gctggccgcc
     4081 agcacgccct cgccccccga ggtggtgtac agcgcccacg cattccacca acctcatgtc
     4141 agctccccgc gcttccagcc cagggctaag agaggttcag gatctggcgg gggctcaggg
     4201 ggaggaaccg gtgctggaag tggtgagggc aggggatctc tccttacttg tggcgacgtg
     4261 gaggagaacc ccggccccat ggaattcggc ctgagctggc tgttcctggt ggccatcctg
     4321 aagggcgtgc agtgcagcag agacatcctg ctgacccaga cccctctgag cctgcctgtg
     4381 tctctgggcg atcaggccag catcagctgc agatccagcc agagcctggt gcaccggaac
     4441 ggcaacacct acctgcactg gtatctgcag aagcccggcc agagccccaa gctgctgatc
     4501 cacaaggtgt ccaaccggtt cagcggcgtg cccgacagat tttctggcag cggctccggc
     4561 accgacttca ccctgaagat cagccgggtg gaagccgagg acctgggcgt gtacttctgc
     4621 agccagtcca cccacgtgcc ccccctgaca tttggcgccg gaacaaagct ggaactgaag
     4681 gggggaggcg gatctggcgg cggaggaagt ggcggagggg gatctgaagt gaagctgcag
     4741 cagtccggcc ccagcctggt ggaacctggc gcctctgtga tgatctcctg caaggccagc
     4801 ggcagctcct tcaccggcta caacatgaac tgggtgcgcc agaacatcgg caagagcctg
     4861 gaatggatcg gcgccatcga cccctactac ggcggcacca gctacaacca gaagttcaag
     4921 ggcagagcca ccctgaccgt ggacaagagc agcagcaccg cctacatgca cctgaagtcc
     4981 ctgaccagcg aggacagcgc cgtgtactac tgcgtgtccg gcatgaagta ctggggccag
     5041 ggcacaagcg tgaccgtgtc tagcgccaag accacccccc ctagcgtgta cggaagagtg
     5101 acagtgtcct ctgccgagcc caagagctgc gacaagaccc acacctgtcc cccttgtcct
     5161 gcccctcctg ttgcaggccc ttccgtgttc ctgttccccc caaagcccaa ggacacactg
     5221 atgatcagca gaacccccga agtgacctgc gtggtggtgg acgtgtccca cgaggaccca
     5281 gaagtgaagt tcaattggta cgtggacggc gtggaagtgc acaacgccaa gacaaagccc
     5341 agagaggaac agtaccaaag cacctaccgg gtggtgtccg tgctgaccgt gctgcatcag
     5401 gattggctga acggcaaaga gtacaagtgc aaagtgtcca acaaggccct gcctgccccc
     5461 atcgagaaaa ccatcagcaa ggccaagggc cagccccgcg aaccccaggt gtacacactg
     5521 ccccctagca gggacgagct gaccaagaac caggtgtccc tgacatgcct cgtgaagggc
     5581 ttctacccct ccgatatcgc cgtggaatgg gagagcaacg gccagcccga gaacaactac
     5641 aagacaaccc ctcccgtgct ggacagcgac ggctcattct tcctgtacag caagctgaca
     5701 gtggataagt cccggtggca gcagggcaac gtgttcagct gctccgtgat gcacgaggcc
     5761 ctgcacaacc actacaccca gaaaagcctg tccctgagcc ccggcaagaa ggaccccaaa
     5821 gcggccgcaa ttgaagttat gtatcctcct ccttacctag acaatgagaa gagcaatgga
     5881 accattatcc atgtgaaagg gaaacacctt tgtccaagtc ccctatttcc cggaccttct
     5941 aagccctttt gggtgctggt ggtggttggg ggagtcctgg cttgctatag cttgctagta
     6001 acagtggcct ttattatttt ctgggtgagg agtaagagga gcaggctcct gcacagtgac
     6061 tacatgaaca tgactccccg ccgccccggg cccacccgca agcattacca gccctatgcc
     6121 ccaccacgcg acttcgcagc ctatcgctcc agagtgaagt tcagcaggag cgcagacgcc
     6181 cccgcgtacc agcagggcca gaaccagctc tataacgagc tcaatctagg acgaagagag
     6241 gagtacgatg ttttggacaa gagacgtggc cgggaccctg agatgggggg aaagccgaga
     6301 aggaagaacc ctcaggaagg cctgtacaat gaactgcaga aagataagat ggcggaggcc
     6361 tacagtgaga ttgggatgaa aggcgagcgc cggaggggca aggggcacga tggcctttac
     6421 cagggtctca gtacagccac caaggacacc tacgacgccc ttcacatgca ggccctgccc
     6481 cctcgctaag gatccgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     6541 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     6601 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
     6661 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     6721 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     6781 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     6841 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     6901 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa
     6961 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     7021 gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa
     7081 caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt
     7141 gaatccacat actccaatac tcctgaaatc catcgatgga gttcattatg gacagcgcag
     7201 aaagagctgg ggagaattgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7261 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7321 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7381 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7441 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7501 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7561 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7621 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7681 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7741 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7801 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7861 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7921 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7981 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     8041 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     8101 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     8161 gcagattacg cgcagaaaaa aaggatctca a
//
