LOCUS       sequence-412347-        8809 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 8809)
  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 8809)
  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-829_HA-GD2-28z_CAR_JUN_Retroviral
FEATURES             Location/Qualifiers
     source          1..8809
                     /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             4843..4896
                     /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             7157..7671
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7843..7859
                     /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(7867..7873,7874..7891,7892..7897))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7912..7933
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8050..8067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8221..8809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8301..8320)
                     /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 gccaccatga ccgcgaaaat ggagacaaca ttttacgatg acgctcttaa tgcgtccttt
     3841 ctgccatcag aatccggccc atacggatat agcaatccga aaatattgaa gcaatctatg
     3901 acattgaatc ttgctgatcc tgtcggttcc ttgaaacctc atctgcgggc aaagaattcc
     3961 gatctgttga ctagcccaga tgtaggactc ctgaaactcg catcccctga actcgaacgg
     4021 ctcattatac aatcaagtaa tggacatatt acgacgaccc ctactcctac gcaatttctt
     4081 tgtcctaaga atgtcaccga cgaacaagaa ggttttgcag aagggtttgt acgggcattg
     4141 gcggagcttc attcacaaaa taccctgcct agtgtgacct ctgccgctca accagtgaat
     4201 ggagctggaa tggtcgcccc tgccgttgca agcgttgccg gcggaagtgg atctggcggg
     4261 tttagtgcat cacttcattc tgaaccacca gtgtatgcca atctgtccaa ttttaatccc
     4321 ggagccctct catctggagg aggcgcccct tcttatggtg cagcgggact tgcgttccct
     4381 gcacagccac aacaacaaca acaaccccct catcatttgc cacaacaaat gcctgtccaa
     4441 catccacgcc tgcaagctct caaagaagaa ccccaaaccg tccctgaaat gccaggggaa
     4501 accccaccac tttcaccgat tgatatggaa tctcaagaaa gaataaaagc cgaaagaaaa
     4561 cgtatgcgca atcggatagc cgcgtcaaaa tgtcgcaagc gcaaactcga acggattgca
     4621 cgcttggaag agaaggttaa gactctgaag gcgcaaaatt ctgaacttgc ttcaaccgct
     4681 aatatgctgc gagagcaagt cgcccaattg aagcaaaagg taatgaatca tgtgaatagc
     4741 ggatgtcagc tgatgctcac tcaacaactg cagactttca gggctaagag aggttcagga
     4801 tctggaggcg gctcgggggg aggaaccggt gctggaagtg gtgagggcag gggatctctc
     4861 cttacttgtg gcgacgtgga ggagaacccc ggccccatgg aattcggcct gagctggctg
     4921 ttcctggtgg ccatcctgaa gggcgtgcag tgcagcagag acatcctgct gacccagacc
     4981 cctctgagcc tgcctgtgtc tctgggcgat caggccagca tcagctgcag atccagccag
     5041 agcctggtgc accggaacgg caacacctac ctgcactggt atctgcagaa gcccggccag
     5101 agccccaagc tgctgatcca caaggtgtcc aaccggttca gcggcgtgcc cgacagattt
     5161 tctggcagcg gctccggcac cgacttcacc ctgaagatca gccgggtgga agccgaggac
     5221 ctgggcgtgt acttctgcag ccagtccacc cacgtgcccc ccctgacatt tggcgccgga
     5281 acaaagctgg aactgaaggg gggaggcgga tctggcggcg gaggaagtgg cggaggggga
     5341 tctgaagtga agctgcagca gtccggcccc agcctggtgg aacctggcgc ctctgtgatg
     5401 atctcctgca aggccagcgg cagctccttc accggctaca acatgaactg ggtgcgccag
     5461 aacatcggca agagcctgga atggatcggc gccatcgacc cctactacgg cggcaccagc
     5521 tacaaccaga agttcaaggg cagagccacc ctgaccgtgg acaagagcag cagcaccgcc
     5581 tacatgcacc tgaagtccct gaccagcgag gacagcgccg tgtactactg cgtgtccggc
     5641 atgaagtact ggggccaggg cacaagcgtg accgtgtcta gcgccaagac caccccccct
     5701 agcgtgtacg gaagagtgac agtgtcctct gccgagccca agagctgcga caagacccac
     5761 acctgtcccc cttgtcctgc ccctcctgtt gcaggccctt ccgtgttcct gttcccccca
     5821 aagcccaagg acacactgat gatcagcaga acccccgaag tgacctgcgt ggtggtggac
     5881 gtgtcccacg aggacccaga agtgaagttc aattggtacg tggacggcgt ggaagtgcac
     5941 aacgccaaga caaagcccag agaggaacag taccaaagca cctaccgggt ggtgtccgtg
     6001 ctgaccgtgc tgcatcagga ttggctgaac ggcaaagagt acaagtgcaa agtgtccaac
     6061 aaggccctgc ctgcccccat cgagaaaacc atcagcaagg ccaagggcca gccccgcgaa
     6121 ccccaggtgt acacactgcc ccctagcagg gacgagctga ccaagaacca ggtgtccctg
     6181 acatgcctcg tgaagggctt ctacccctcc gatatcgccg tggaatggga gagcaacggc
     6241 cagcccgaga acaactacaa gacaacccct cccgtgctgg acagcgacgg ctcattcttc
     6301 ctgtacagca agctgacagt ggataagtcc cggtggcagc agggcaacgt gttcagctgc
     6361 tccgtgatgc acgaggccct gcacaaccac tacacccaga aaagcctgtc cctgagcccc
     6421 ggcaagaagg accccaaagc ggccgcaatt gaagttatgt atcctcctcc ttacctagac
     6481 aatgagaaga gcaatggaac cattatccat gtgaaaggga aacacctttg tccaagtccc
     6541 ctatttcccg gaccttctaa gcccttttgg gtgctggtgg tggttggggg agtcctggct
     6601 tgctatagct tgctagtaac agtggccttt attattttct gggtgaggag taagaggagc
     6661 aggctcctgc acagtgacta catgaacatg actccccgcc gccccgggcc cacccgcaag
     6721 cattaccagc cctatgcccc accacgcgac ttcgcagcct atcgctccag agtgaagttc
     6781 agcaggagcg cagacgcccc cgcgtaccag cagggccaga accagctcta taacgagctc
     6841 aatctaggac gaagagagga gtacgatgtt ttggacaaga gacgtggccg ggaccctgag
     6901 atggggggaa agccgagaag gaagaaccct caggaaggcc tgtacaatga actgcagaaa
     6961 gataagatgg cggaggccta cagtgagatt gggatgaaag gcgagcgccg gaggggcaag
     7021 gggcacgatg gcctttacca gggtctcagt acagccacca aggacaccta cgacgccctt
     7081 cacatgcagg ccctgccccc tcgctaagga tccgataaaa taaaagattt tatttagtct
     7141 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
     7201 gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta
     7261 ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     7321 ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag
     7381 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     7441 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     7501 aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg
     7561 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     7621 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tgggtaacag
     7681 tttcttgaag ttggagaaca acattctgag ggtaggagtc gaatattaag taatcctgac
     7741 tcaattagcc actgttttga atccacatac tccaatactc ctgaaatcca tcgatggagt
     7801 tcattatgga cagcgcagaa agagctgggg agaattgtga aattgttatc cgctcacaat
     7861 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     7921 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     7981 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     8041 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     8101 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     8161 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     8221 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     8281 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     8341 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     8401 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     8461 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     8521 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     8581 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     8641 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     8701 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     8761 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaa
//
