LOCUS       sequence-412355-        8830 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 8830)
  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 8830)
  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-831_HA-GD2-28z_CAR_TFAP4_Retroviral
FEATURES             Location/Qualifiers
     source          1..8830
                     /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"
     regulatory      3781..3790
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4864..4917
                     /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             7178..7692
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7864..7880
                     /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(7888..7894,7895..7912,7913..7918))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7933..7954
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8071..8088)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8242..8830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8322..8341)
                     /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 gccaccatgg agtatttcat ggtgcccact cagaaggtgc cctctttgca acatttcagg
     3841 aaaacagaga aagaagtgat aggagggctc tgtagccttg ccaacattcc actaaccccc
     3901 gagactcagc gggaccagga gcggcggatt cggcgggaga tcgccaacag caacgagcgg
     3961 agacgcatgc agagcatcaa cgcgggattc cagtccctca agaccctcat cccccacaca
     4021 gacggagaga agctcagcaa ggcagccatt ctccagcaga cagccgagta catcttctcc
     4081 ctggagcagg agaagaccag gctcttgcag cagaacacac agctcaagcg cttcatccag
     4141 gagctgagcg gctcgtcccc caagcgacgg cgggcagagg acaaggacga aggcataggc
     4201 tccccggaca tctgggagga cgagaaggcg gaggacctgc ggcgggagat gattgagctg
     4261 cggcagcagc tggacaagga gcgctcggtg cgcatgatgc tggaggagca ggtgcgctcg
     4321 ctggaggccc acatgtaccc ggaaaagctc aaggtgattg cgcagcaggt gcagctgcag
     4381 cagcagcagg aacaggtgag gctgctgcac caggagaagc tggagcggga acagcagcag
     4441 ctgcggaccc agcttctgcc ccctccggcc cccacccacc accccacggt gatcgtgcca
     4501 gcaccgcctc ctcctccctc ccaccacatc aatgtcgtca ccatgggccc ctcctcggtc
     4561 atcaactctg tttccacatc ccggcaaaat ctggacacca tcgtgcaggc aatccagcac
     4621 atcgagggca cccaggaaaa gcaggagctg gaggaggagc agcggcgagc tgtcatcgtg
     4681 aagcctgtcc gcagctgccc ggaggccccc acctctgaca ccgcctccga ctccgaggcc
     4741 tcagacagtg acgccatgga ccagagccgg gaggagccgt cgggggacgg ggagcttccc
     4801 agggctaaga gaggttcagg atctggaggc ggatctgggg gaggaaccgg tgctggaagt
     4861 ggtgagggca ggggatctct ccttacttgt ggcgacgtgg aggagaaccc cggccccatg
     4921 gaattcggcc tgagctggct gttcctggtg gccatcctga agggcgtgca gtgcagcaga
     4981 gacatcctgc tgacccagac ccctctgagc ctgcctgtgt ctctgggcga tcaggccagc
     5041 atcagctgca gatccagcca gagcctggtg caccggaacg gcaacaccta cctgcactgg
     5101 tatctgcaga agcccggcca gagccccaag ctgctgatcc acaaggtgtc caaccggttc
     5161 agcggcgtgc ccgacagatt ttctggcagc ggctccggca ccgacttcac cctgaagatc
     5221 agccgggtgg aagccgagga cctgggcgtg tacttctgca gccagtccac ccacgtgccc
     5281 cccctgacat ttggcgccgg aacaaagctg gaactgaagg ggggaggcgg atctggcggc
     5341 ggaggaagtg gcggaggggg atctgaagtg aagctgcagc agtccggccc cagcctggtg
     5401 gaacctggcg cctctgtgat gatctcctgc aaggccagcg gcagctcctt caccggctac
     5461 aacatgaact gggtgcgcca gaacatcggc aagagcctgg aatggatcgg cgccatcgac
     5521 ccctactacg gcggcaccag ctacaaccag aagttcaagg gcagagccac cctgaccgtg
     5581 gacaagagca gcagcaccgc ctacatgcac ctgaagtccc tgaccagcga ggacagcgcc
     5641 gtgtactact gcgtgtccgg catgaagtac tggggccagg gcacaagcgt gaccgtgtct
     5701 agcgccaaga ccaccccccc tagcgtgtac ggaagagtga cagtgtcctc tgccgagccc
     5761 aagagctgcg acaagaccca cacctgtccc ccttgtcctg cccctcctgt tgcaggccct
     5821 tccgtgttcc tgttcccccc aaagcccaag gacacactga tgatcagcag aacccccgaa
     5881 gtgacctgcg tggtggtgga cgtgtcccac gaggacccag aagtgaagtt caattggtac
     5941 gtggacggcg tggaagtgca caacgccaag acaaagccca gagaggaaca gtaccaaagc
     6001 acctaccggg tggtgtccgt gctgaccgtg ctgcatcagg attggctgaa cggcaaagag
     6061 tacaagtgca aagtgtccaa caaggccctg cctgccccca tcgagaaaac catcagcaag
     6121 gccaagggcc agccccgcga accccaggtg tacacactgc cccctagcag ggacgagctg
     6181 accaagaacc aggtgtccct gacatgcctc gtgaagggct tctacccctc cgatatcgcc
     6241 gtggaatggg agagcaacgg ccagcccgag aacaactaca agacaacccc tcccgtgctg
     6301 gacagcgacg gctcattctt cctgtacagc aagctgacag tggataagtc ccggtggcag
     6361 cagggcaacg tgttcagctg ctccgtgatg cacgaggccc tgcacaacca ctacacccag
     6421 aaaagcctgt ccctgagccc cggcaagaag gaccccaaag cggccgcaat tgaagttatg
     6481 tatcctcctc cttacctaga caatgagaag agcaatggaa ccattatcca tgtgaaaggg
     6541 aaacaccttt gtccaagtcc cctatttccc ggaccttcta agcccttttg ggtgctggtg
     6601 gtggttgggg gagtcctggc ttgctatagc ttgctagtaa cagtggcctt tattattttc
     6661 tgggtgagga gtaagaggag caggctcctg cacagtgact acatgaacat gactccccgc
     6721 cgccccgggc ccacccgcaa gcattaccag ccctatgccc caccacgcga cttcgcagcc
     6781 tatcgctcca gagtgaagtt cagcaggagc gcagacgccc ccgcgtacca gcagggccag
     6841 aaccagctct ataacgagct caatctagga cgaagagagg agtacgatgt tttggacaag
     6901 agacgtggcc gggaccctga gatgggggga aagccgagaa ggaagaaccc tcaggaaggc
     6961 ctgtacaatg aactgcagaa agataagatg gcggaggcct acagtgagat tgggatgaaa
     7021 ggcgagcgcc ggaggggcaa ggggcacgat ggcctttacc agggtctcag tacagccacc
     7081 aaggacacct acgacgccct tcacatgcag gccctgcccc ctcgctaagg atccgataaa
     7141 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     7201 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata actgagaata
     7261 gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca aacaggatat
     7321 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
     7381 cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     7441 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     7501 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc
     7561 cgatagactg cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     7621 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     7681 ggggtctttc atgggtaaca gtttcttgaa gttggagaac aacattctga gggtaggagt
     7741 cgaatattaa gtaatcctga ctcaattagc cactgttttg aatccacata ctccaatact
     7801 cctgaaatcc atcgatggag ttcattatgg acagcgcaga aagagctggg gagaattgtg
     7861 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     7921 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7981 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     8041 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     8101 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     8161 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     8221 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     8281 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     8341 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     8401 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     8461 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     8521 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     8581 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     8641 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     8701 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     8761 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     8821 aggatctcaa
//
