LOCUS       sequence-141876-        9085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral transfer vector which encodes the Disialoganglioside 
            (GD2) mini-operon. Mammalian cells transduced with this vector 
            become GD2 positive..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9085)
  AUTHORS   Thomas S, Straathof K, Himoudi N, Anderson J, Pule M
  TITLE     An Optimized GD2-Targeting Retroviral Cassette for More Potent and 
            Safer Cellular Therapy of Neuroblastoma and Other Cancers.
  JOURNAL   PLoS One. 2016 Mar 31;11(3):e0152196. doi: 
            10.1371/journal.pone.0152196. eCollection 2016.
  PUBMED    27030986
REFERENCE   2  (bases 1 to 9085)
  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"; journalName: "PLoS One.";
            date: "2016-03-31"; volume: "11(3):e0152196. doi"; pages: " 
            10.1371/journal.pone.0152196. eCollection 2016"
COMMENT     Sequence Label: MP9956:SFG.GD3synthase-2A-GD2synthase
FEATURES             Location/Qualifiers
     source          1..9085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    655..1012
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     primer_bind     1423..1445
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1461..1477
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1503..1877
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1815..1831
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             2948..3007
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     LTR             4890..5359
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6058..6075)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6058..6074)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6067..6089)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6283..6302)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6402..6424
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6463..6481)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6549..6653
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6654..6722,6723..7514)
                     /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     complement(6872..6891)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7685..8273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8174..8193
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8427..8444
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8561..8582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8597..8602,8603..8620,8621..8627)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8635..8651
                     /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)."
     primer_bind     8640..8662
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8659..8675
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8659..8675
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     LTR             join(9084..9085,1..592)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca gatggaacag
      121 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      241 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      301 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      361 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttatgctccc cgagctcaat
      421 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      481 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      541 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      601 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg
      661 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      721 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      781 gttcggaaca cccggccgca accctgggag acgtcccagg gacttcgggg gccgtttttg
      841 tggcccgacc tgagtcctaa aatcccgatc gtttaggact ctttggtgca ccccccttag
      901 aggagggata tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg
      961 aatttttgct ttcggtttgg gaccgaagcc gcgccgcgcg tcttgtctgc tgcagcatcg
     1021 ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaatatg ggcccgggct
     1081 agcctgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     1141 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     1201 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     1261 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtggggtaca
     1321 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     1381 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     1441 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gcccccatat
     1501 ggccatatga gatcttatat ggggcacccc cgccccttgt aaacttccct gaccctgaca
     1561 tgacaagagt tactaacagc ccctctctcc aagctcactt acaggctctc tacttagtcc
     1621 agcacgaagt ctggagacct ctggcggcag cctaccaaga acaactggac cgaccggtgg
     1681 tacctcaccc ttaccgagtc ggcgacacag tgtgggtccg ccgacaccag actaagaacc
     1741 tagaacctcg ctggaaagga ccttacacag tcctgctgac cacccccacc gccctcaaag
     1801 tagacggcat cgcagcttgg atacacgccg cccacgtgaa ggctgccgac cccgggggtg
     1861 gaccatcctc tagactgcca ccatgagccc ctgcgggcgg gccctacata cgtccagagg
     1921 ggccatggcc atgctagctc ggaaattccc gcgtacccgg ctacccgtag gagccagtgc
     1981 cctctgtgtg gtggtcctct gttggctcta catcttccct gtctaccggc tgcccaacga
     2041 gaaggagatt gtgcaagggg tgctggcaca gcggacggct tggaggacga accagacgtc
     2101 agccagtctc ttcaggagac aaatggaaga ctgctgtgac cctgcccatc tctttgctat
     2161 gactaagatg aactccccca tggggaagag cctgtggtac gatggggagc tcttatactc
     2221 gttcaccatc gataattcca cgtactccct cttcccccag gcaaccccct tccagctgcc
     2281 attgaagaaa tgtgcggtgg tgggaaacgg tgggattctg aagatgagtg gctgtggccg
     2341 tcaaatagat gaagcaaatt ttgtcatgcg gtgcaacctg cctcccttgt caagtgagta
     2401 caccagagat gtcggttcca aaactcagtt ggtgacagct aaccccagca taattcgcca
     2461 gagatttgaa aacctgctgt ggtccagaaa gaagtttgtg gacaacatga agatctataa
     2521 ccacagttac atctacatgc ctgccttttc tatgaagaca ggcacagagc cgtctctccg
     2581 tgtgtattac acactgaaag atgttggagc caatcaaaca gtgctttttg ctaaccccaa
     2641 ctttcttcgg aacattggaa agttctggaa gagtaggggg atccacgcca agcgcttgtc
     2701 tacaggactc tttttggtga gtgcagcctt gggcctctgt gaggaggtgt ccatctatgg
     2761 cttctggccc ttctcggtga acatgcaagg ggaccctatc agtcaccact actatgacaa
     2821 cgtcttgccc ttctcaggct accatgccat gcctgaggaa ttccttcagc tttggtatct
     2881 tcacaaaatc ggcgcgctga ggatgcagct ggacccgtgt gaggagccgt caccacagcc
     2941 cacttcccag tgcaccaact acgccctgct gaagctggcc ggcgacgtgg agagcaaccc
     3001 cggccccatg cggctagacc gccgggccct ctatgcgcta gtcttgctgc tcgcctgcgc
     3061 ctcgctgggt ctcctgtact ccagcacccg aaacgcgcca agtctcccga accctctggc
     3121 attgtggtcg cccccacaag gtcccccgag actcgatctg ctagaccttg cccctgagcc
     3181 tcgctacgca cacatcccgg tcaggatcaa ggagcaagtg gtggggctgc tggctcagaa
     3241 caactgcagt tgtgaatcca agggaggaag ccttcccttg ccctttctga gacaggttcg
     3301 ggcggttgac ctcactaaag cctttgacgc tgaggagctg agggctgttt ctgtcgccag
     3361 ggagcaggaa taccaggcct tccttgcaag gagccggtcc ctggctgacc agctgctgat
     3421 agctcccgcc aactccccct tacagtaccc cctgcagggt gtggaggttc agcccctcag
     3481 gagcatcctg gtgccagggc taagtctgca ggaagcttct gtccaggaga tataccaggt
     3541 gaacctgagt gcttctctag gcacctggga cgtggcaggg gaagtaacag gagtgactct
     3601 cacgggagag gggcaaccgg atctcaccct tgccagccca gttctggata aactcaaccg
     3661 gcagctgcaa ctggtgactt acagcagccg gagctaccag gccaacacag cagacacagt
     3721 ccggttctcc accaagggac atgaagttgc cttcaccatc ctcgtaagac accctcccaa
     3781 cccccggctg tacccaccat catccctacc ccaaggagcc gagtacaaca tcagtgctct
     3841 ggttaccatt gccaccaaga cttttcttcg ttatgatcga ctacggacac tcattgccag
     3901 catccgacgc ttttacccta cggtcaccat agtcatcgct gacgacagtg acaaaccgga
     3961 gcgcattagc gacccacacg tggagcacta cttcatgccc ttcggcaagg gttggttcgc
     4021 aggtcggaac ctggcggtgt cccaagtaac caccaaatac gtgctgtggg tggacgacga
     4081 ctttgtcttc acggcgcgca cgcggctgga gaagcttgtg gacgtgctcg agaagacgcc
     4141 cctggacctg gtagggggcg cggtgcggga gatctcgggc tacgctacca cctaccggca
     4201 gctgctgagt gtggagccgg gcgccccagg ccttgggaac tgcttccggc aaaagcaggg
     4261 cttccaccac gagctcgttg gcttcccaag ttgtgtggtc acagacggcg tggtcaactt
     4321 cttcctggcg cgcacagaca aagtgcgcca agtgggcttt gacccacgcc tcaaccgggt
     4381 ggcacatctg gaattcttcc tggatggact tggtttcctt cgtgttggct cctgctctga
     4441 tgttgttgtg gatcatgcat cgaaggtgaa gctgccgtgg acagcaaaag acccaggagc
     4501 tgaaacttat gcccgttaca ggtacccggg atcactggac caaagtcaag tggccaaaca
     4561 tcgcctactc ttcttcaaac accggctaca gtgcatgacc gccgagtgaa cgcgtcatca
     4621 tcgatccgga ttagtccaat ttgttaaaga caggatatca gtggtccagg ctctagtttt
     4681 gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat
     4741 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     4801 agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt
     4861 cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     4921 aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa
     4981 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
     5041 gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
     5101 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     5161 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc
     5221 cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
     5281 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
     5341 cgtcagcggg ggtctttcac acatgcagca tgtatcaaaa ttaatttggt tttttttctt
     5401 aagtatttac attaaatggc catagtactt aaagttacat tggcttcctt gaaataaaca
     5461 tggagtattc agaatgtgtc ataaatattt ctaattttaa gatagtatct ccattggctt
     5521 tctacttttt cttttatttt tttttgtcct ctgtcttcca tttgttgttg ttgttgtttg
     5581 tttgtttgtt tgttggttgg ttggttaatt tttttttaaa gatcctacac tatagttcaa
     5641 gctagactat tagctactct gtaacccagg gtgaccttga agtcatgggt agcctgctgt
     5701 tttagccttc ccacatctaa gattacaggt atgagctatc atttttggta tattgattga
     5761 ttgattgatt gatgtgtgtg tgtgtgattg tgtttgtgtg tgtgactgtg aaaatgtgtg
     5821 tatgggtgtg tgtgaatgtg tgtatgtatg tgtgtgtgtg agtgtgtgtg tgtgtgtgtg
     5881 catgtgtgtg tgtgtgactg tgtctatgtg tatgactgtg tgtgtgtgtg tgtgtgtgtg
     5941 tgtgtgtgtg tgtgtgtgtg tgttgtgaaa aaatattcta tggtagtgag agccaacgct
     6001 ccggctcagg tgtcaggttg gtttttgaga cagagtcttt cacttagctt ggaattcact
     6061 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     6121 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     6181 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     6241 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     6301 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     6361 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     6421 gaggttttca ccgtcatcac cgaaacgcgc gatgacgaaa gggcctcgtg atacgcctat
     6481 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     6541 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6601 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     6661 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6721 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     6781 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6841 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     6901 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     6961 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     7021 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     7081 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     7141 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     7201 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     7261 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     7321 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     7381 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     7441 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     7501 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     7561 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7621 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7681 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7741 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7801 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     7861 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7921 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7981 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     8041 cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc acgcttcccg
     8101 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     8161 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     8221 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     8281 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     8341 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     8401 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     8461 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     8521 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     8581 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     8641 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctttgctctt
     8701 aggagtttcc taatacatcc caaactcaaa tatataaagc atttgacttg ttctatgccc
     8761 tagggggcgg ggggaagcta agccagcttt ttttaacatt taaaatgtta attccatttt
     8821 aaatgcacag atgtttttat ttcataaggg tttcaatgtg catgaatgct gcaatattcc
     8881 tgttaccaaa gctagtataa ataaaaatag ataaacgtgg aaattactta gagtttctgt
     8941 cattaacgtt tccttcctca gttgacaaca taaatgcgct gctgagcaag ccagtttgca
     9001 tctgtcagga tcaatttccc attatgccag tcatattaat tactagtcaa ttagttgatt
     9061 tttatttttg acatatacat gtgaa
//
