LOCUS       sequence-412352-        8647 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 8647)
  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 8647)
  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-830_HA-GD2-28z_CAR_tNGFR_Retroviral
FEATURES             Location/Qualifiers
     source          1..8647
                     /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             4681..4734
                     /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             6995..7509
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7681..7697
                     /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(7705..7711,7712..7729,7730..7735))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7750..7771
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7888..7905)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8059..8647)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8139..8158)
                     /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 gcgctggtgc aactggacgg gccatggatg gaccacggct tttgctcctg
     3841 ttgttgttgg gcgtcagcct tggaggggct aaagaggcct gcccgaccgg tttgtacaca
     3901 cactccggcg agtgttgtaa ggcgtgtaat cttggcgaag gggtcgccca accgtgtggc
     3961 gccaaccaga cggtttgtga gccttgcttg gatagcgtca cgttttccga cgtcgtttca
     4021 gccaccgaac catgcaaacc ttgtacggaa tgcgtgggtc tgcagagcat gagcgcaccg
     4081 tgcgtagagg cggacgatgc ggtatgccga tgtgcctacg gctactacca agacgagacg
     4141 acagggaggt gtgaggcatg cagagtatgt gaggccgggt ctggactcgt attttcctgt
     4201 caagacaaac aaaacacagt atgtgaggag tgcccggatg ggacttattc cgatgaagcg
     4261 aatcatgtgg acccgtgctt gccatgtacg gtctgtgagg ataccgaaag acagctccga
     4321 gaatgcaccc gctgggctga cgcggaatgc gaagaaatcc caggtcgatg gatcacccgc
     4381 tcaacccccc ccgaagggtc agattctacg gcgccctcta ctcaggagcc agaagctcca
     4441 cccgaacaag acctgatcgc ttctacggtg gcaggtgtcg ttacaacagt catgggttcc
     4501 tcacaacctg tggttacgcg ggggacaact gacaatttga ttccagtata ttgtagcatt
     4561 ctggccgcgg tcgtggtagg gcttgtcgcg tatatagcgt tcaaaaggtg gaacagcagg
     4621 gctaagagag gttcaggatc tggggggggc tcgggaggtg gaaccggtgc tggaagtggt
     4681 gagggcaggg gatctctcct tacttgtggc gacgtggagg agaaccccgg ccccatggaa
     4741 ttcggcctga gctggctgtt cctggtggcc atcctgaagg gcgtgcagtg cagcagagac
     4801 atcctgctga cccagacccc tctgagcctg cctgtgtctc tgggcgatca ggccagcatc
     4861 agctgcagat ccagccagag cctggtgcac cggaacggca acacctacct gcactggtat
     4921 ctgcagaagc ccggccagag ccccaagctg ctgatccaca aggtgtccaa ccggttcagc
     4981 ggcgtgcccg acagattttc tggcagcggc tccggcaccg acttcaccct gaagatcagc
     5041 cgggtggaag ccgaggacct gggcgtgtac ttctgcagcc agtccaccca cgtgcccccc
     5101 ctgacatttg gcgccggaac aaagctggaa ctgaaggggg gaggcggatc tggcggcgga
     5161 ggaagtggcg gagggggatc tgaagtgaag ctgcagcagt ccggccccag cctggtggaa
     5221 cctggcgcct ctgtgatgat ctcctgcaag gccagcggca gctccttcac cggctacaac
     5281 atgaactggg tgcgccagaa catcggcaag agcctggaat ggatcggcgc catcgacccc
     5341 tactacggcg gcaccagcta caaccagaag ttcaagggca gagccaccct gaccgtggac
     5401 aagagcagca gcaccgccta catgcacctg aagtccctga ccagcgagga cagcgccgtg
     5461 tactactgcg tgtccggcat gaagtactgg ggccagggca caagcgtgac cgtgtctagc
     5521 gccaagacca ccccccctag cgtgtacgga agagtgacag tgtcctctgc cgagcccaag
     5581 agctgcgaca agacccacac ctgtccccct tgtcctgccc ctcctgttgc aggcccttcc
     5641 gtgttcctgt tccccccaaa gcccaaggac acactgatga tcagcagaac ccccgaagtg
     5701 acctgcgtgg tggtggacgt gtcccacgag gacccagaag tgaagttcaa ttggtacgtg
     5761 gacggcgtgg aagtgcacaa cgccaagaca aagcccagag aggaacagta ccaaagcacc
     5821 taccgggtgg tgtccgtgct gaccgtgctg catcaggatt ggctgaacgg caaagagtac
     5881 aagtgcaaag tgtccaacaa ggccctgcct gcccccatcg agaaaaccat cagcaaggcc
     5941 aagggccagc cccgcgaacc ccaggtgtac acactgcccc ctagcaggga cgagctgacc
     6001 aagaaccagg tgtccctgac atgcctcgtg aagggcttct acccctccga tatcgccgtg
     6061 gaatgggaga gcaacggcca gcccgagaac aactacaaga caacccctcc cgtgctggac
     6121 agcgacggct cattcttcct gtacagcaag ctgacagtgg ataagtcccg gtggcagcag
     6181 ggcaacgtgt tcagctgctc cgtgatgcac gaggccctgc acaaccacta cacccagaaa
     6241 agcctgtccc tgagccccgg caagaaggac cccaaagcgg ccgcaattga agttatgtat
     6301 cctcctcctt acctagacaa tgagaagagc aatggaacca ttatccatgt gaaagggaaa
     6361 cacctttgtc caagtcccct atttcccgga ccttctaagc ccttttgggt gctggtggtg
     6421 gttgggggag tcctggcttg ctatagcttg ctagtaacag tggcctttat tattttctgg
     6481 gtgaggagta agaggagcag gctcctgcac agtgactaca tgaacatgac tccccgccgc
     6541 cccgggccca cccgcaagca ttaccagccc tatgccccac cacgcgactt cgcagcctat
     6601 cgctccagag tgaagttcag caggagcgca gacgcccccg cgtaccagca gggccagaac
     6661 cagctctata acgagctcaa tctaggacga agagaggagt acgatgtttt ggacaagaga
     6721 cgtggccggg accctgagat ggggggaaag ccgagaagga agaaccctca ggaaggcctg
     6781 tacaatgaac tgcagaaaga taagatggcg gaggcctaca gtgagattgg gatgaaaggc
     6841 gagcgccgga ggggcaaggg gcacgatggc ctttaccagg gtctcagtac agccaccaag
     6901 gacacctacg acgcccttca catgcaggcc ctgccccctc gctaaggatc cgataaaata
     6961 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     7021 caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag
     7081 aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg
     7141 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc
     7201 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     7261 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     7321 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga
     7381 tagactgcgt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact
     7441 tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     7501 gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga
     7561 atattaagta atcctgactc aattagccac tgttttgaat ccacatactc caatactcct
     7621 gaaatccatc gatggagttc attatggaca gcgcagaaag agctggggag aattgtgaaa
     7681 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7741 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7801 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7861 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7921 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7981 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     8041 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     8101 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     8161 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8221 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     8281 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8341 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8401 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8461 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     8521 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8581 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8641 atctcaa
//
