LOCUS       sequence-10336-e        8191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8191)
  AUTHORS   Brewin J, Mancao C, Straathof K, Karlsson H, Samarasinghe S, Amrolia
            PJ, Pule M
  TITLE     Generation of EBV-specific cytotoxic T cells that are resistant to 
            calcineurin inhibitors for the treatment of posttransplantation 
            lymphoproliferative disease.
  JOURNAL   Blood. 2009 Nov 26;114(23):4792-803. doi: 
            10.1182/blood-2009-07-228387. Epub 2009 Sep 21.
  PUBMED    19770360
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: 
            "10.1182/blood-2009-07-228387"; journalName: "Blood"; date: 
            "2009-11-26- 26"; volume: "114"; issue: "23"; pages: "4792-803"
COMMENT     Sequence Label: SFG.wtCNb_opt.IRES.eGFP
FEATURES             Location/Qualifiers
     source          1..8191
                     /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"
     regulatory      1877..1886
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2460..3011
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2604..2621)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2831..2850
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3011..3013,3014..3016,3017..3730)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3011..3013,3014..3016,3017..3727)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3056..3077)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3317..3336)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3664..3685
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3996..4465
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5164..5181)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5164..5180)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5173..5195)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5389..5408)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5508..5530
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5569..5587)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5655..5759
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5760..5828,5829..6620)
                     /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(5978..5997)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6791..7379
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7280..7299
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7533..7550
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7667..7688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7703..7708,7709..7726,7727..7733)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7741..7757
                     /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     7746..7768
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7765..7781
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7765..7781
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     LTR             join(8190..8191,1..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 ccatgggcaa cgaggccagc taccctctgg agatgtgctc
     1921 ccacttcgac gccgacgaga tcaagcggct gggcaagcgc ttcaagaagc tggacctgga
     1981 caacagcggc agcctgagcg tggaggagtt tatgtctctg cccgagctgc agcagaaccc
     2041 cctggtgcag cgcgtgatcg acatcttcga caccgacggc aacggcgagg tggacttcaa
     2101 ggagttcatc gagggcgtga gccagttcag cgtgaagggc gacaaggagc agaagctgcg
     2161 gttcgccttc cggatctacg atatggataa agatggctat atttctaatg gcgagctgtt
     2221 ccaggtgctg aagatgatgg tgggcaacaa tctgaaagat acccagctgc agcagatcgt
     2281 ggacaagacc atcatcaacg ccgacaagga cggcgacggc agaatcagct tcgaggagtt
     2341 ctgtgccgtg gtgggaggcc tggatattca caaaaaaatg gtggtggacg tgtgaacgcg
     2401 tacgtcgcga ccgcggacat gtacagagct cgagcgggat caattccgcc ccccccctaa
     2461 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     2521 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     2581 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     2641 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     2701 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     2761 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     2821 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     2881 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     2941 gaggttaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     3001 cgataataat atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     3061 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     3121 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     3181 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     3241 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     3301 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     3361 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     3421 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     3481 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     3541 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3601 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     3661 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3721 gtacaagtaa tccggattag tccaatttgt taaagacagg atatcagtgg tccaggctct
     3781 agttttgact caacaatatc accagctgaa gcctatagag tacgagccat agataaaata
     3841 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     3901 caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga
     3961 gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc
     4021 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg
     4081 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     4141 tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     4201 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     4261 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     4321 gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct
     4381 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     4441 actacccgtc agcgggggtc tttcacacat gcagcatgta tcaaaattaa tttggttttt
     4501 tttcttaagt atttacatta aatggccata gtacttaaag ttacattggc ttccttgaaa
     4561 taaacatgga gtattcagaa tgtgtcataa atatttctaa ttttaagata gtatctccat
     4621 tggctttcta ctttttcttt tatttttttt tgtcctctgt cttccatttg ttgttgttgt
     4681 tgtttgtttg tttgtttgtt ggttggttgg ttaatttttt tttaaagatc ctacactata
     4741 gttcaagcta gactattagc tactctgtaa cccagggtga ccttgaagtc atgggtagcc
     4801 tgctgtttta gccttcccac atctaagatt acaggtatga gctatcattt ttggtatatt
     4861 gattgattga ttgattgatg tgtgtgtgtg tgattgtgtt tgtgtgtgtg actgtgaaaa
     4921 tgtgtgtatg ggtgtgtgtg aatgtgtgta tgtatgtgtg tgtgtgagtg tgtgtgtgtg
     4981 tgtgtgcatg tgtgtgtgtg tgactgtgtc tatgtgtatg actgtgtgtg tgtgtgtgtg
     5041 tgtgtgtgtg tgtgtgtgtg tgtgtgtgtt gtgaaaaaat attctatggt agtgagagcc
     5101 aacgctccgg ctcaggtgtc aggttggttt ttgagacaga gtctttcact tagcttggaa
     5161 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     5221 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     5281 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
     5341 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     5401 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     5461 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5521 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgatg acgaaagggc ctcgtgatac
     5581 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5641 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5701 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5761 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5821 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5881 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5941 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6001 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6061 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6121 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6181 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6241 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6301 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6361 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6421 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6481 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6541 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6601 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6661 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6721 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6781 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6841 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6901 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     6961 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7021 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7081 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7141 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc
     7201 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7261 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7321 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7381 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     7441 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     7501 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     7561 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     7621 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     7681 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     7741 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttt
     7801 gctcttagga gtttcctaat acatcccaaa ctcaaatata taaagcattt gacttgttct
     7861 atgccctagg gggcgggggg aagctaagcc agcttttttt aacatttaaa atgttaattc
     7921 cattttaaat gcacagatgt ttttatttca taagggtttc aatgtgcatg aatgctgcaa
     7981 tattcctgtt accaaagcta gtataaataa aaatagataa acgtggaaat tacttagagt
     8041 ttctgtcatt aacgtttcct tcctcagttg acaacataaa tgcgctgctg agcaagccag
     8101 tttgcatctg tcaggatcaa tttcccatta tgccagtcat attaattact agtcaattag
     8161 ttgattttta tttttgacat atacatgtga a
//
