LOCUS       sequence-10337-e        8197 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 8197)
  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 8197)
  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.CNb30_opt.IRES.eGFP
FEATURES             Location/Qualifiers
     source          1..8197
                     /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    2466..3017
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2610..2627)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2837..2856
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3017..3019,3020..3022,3023..3736)
                     /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(3017..3019,3020..3022,3023..3733)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3062..3083)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3323..3342)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3670..3691
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4002..4471
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5170..5187)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5170..5186)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5179..5201)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5395..5414)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5514..5536
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5575..5593)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5661..5765
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5766..5834,5835..6626)
                     /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(5984..6003)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6797..7385
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7286..7305
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7539..7556
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7673..7694
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7709..7714,7715..7732,7733..7739)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7747..7763
                     /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     7752..7774
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7771..7787
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7771..7787
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     LTR             join(8196..8197,1..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 taggctgcca 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 taccaagctg gccgataccc agctgcagca
     2281 gatcgtggac aagaccatca tcaacgccga caaggacggc gacggcagaa tcagcttcga
     2341 ggagttctgt gccgtggtgg gaggcctgga tattcacaaa aaaatggtgg tggacgtgtg
     2401 aacgcgtacg tcgcgaccgc ggacatgtac agagctcgag cgggatcaat tccgcccccc
     2461 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     2521 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     2581 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     2641 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     2701 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     2761 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     2821 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     2881 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     2941 ttagtcgagg ttaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
     3001 aaaacacgat aataatatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3061 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     3121 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3181 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     3241 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3301 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3361 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     3421 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     3481 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     3541 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3601 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     3661 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3721 cgagctgtac aagtaatccg gattagtcca atttgttaaa gacaggatat cagtggtcca
     3781 ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat
     3841 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     3901 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag
     3961 aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag
     4021 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
     4081 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     4141 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
     4201 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     4261 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     4321 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     4381 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     4441 tgattgacta cccgtcagcg ggggtctttc acacatgcag catgtatcaa aattaatttg
     4501 gttttttttc ttaagtattt acattaaatg gccatagtac ttaaagttac attggcttcc
     4561 ttgaaataaa catggagtat tcagaatgtg tcataaatat ttctaatttt aagatagtat
     4621 ctccattggc tttctacttt ttcttttatt tttttttgtc ctctgtcttc catttgttgt
     4681 tgttgttgtt tgtttgtttg tttgttggtt ggttggttaa ttttttttta aagatcctac
     4741 actatagttc aagctagact attagctact ctgtaaccca gggtgacctt gaagtcatgg
     4801 gtagcctgct gttttagcct tcccacatct aagattacag gtatgagcta tcatttttgg
     4861 tatattgatt gattgattga ttgatgtgtg tgtgtgtgat tgtgtttgtg tgtgtgactg
     4921 tgaaaatgtg tgtatgggtg tgtgtgaatg tgtgtatgta tgtgtgtgtg tgagtgtgtg
     4981 tgtgtgtgtg tgcatgtgtg tgtgtgtgac tgtgtctatg tgtatgactg tgtgtgtgtg
     5041 tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgttgtga aaaaatattc tatggtagtg
     5101 agagccaacg ctccggctca ggtgtcaggt tggtttttga gacagagtct ttcacttagc
     5161 ttggaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     5221 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     5281 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
     5341 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
     5401 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     5461 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5521 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgatgacga aagggcctcg
     5581 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5641 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5701 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5761 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5821 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5881 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5941 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6001 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6061 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6121 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6181 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6241 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6301 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6361 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6421 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6481 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6541 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6601 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6661 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6721 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6781 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6841 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6901 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6961 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7021 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7081 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7141 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg
     7201 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7261 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7321 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7381 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7441 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     7501 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     7561 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     7621 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     7681 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     7741 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     7801 agctttgctc ttaggagttt cctaatacat cccaaactca aatatataaa gcatttgact
     7861 tgttctatgc cctagggggc ggggggaagc taagccagct ttttttaaca tttaaaatgt
     7921 taattccatt ttaaatgcac agatgttttt atttcataag ggtttcaatg tgcatgaatg
     7981 ctgcaatatt cctgttacca aagctagtat aaataaaaat agataaacgt ggaaattact
     8041 tagagtttct gtcattaacg tttccttcct cagttgacaa cataaatgcg ctgctgagca
     8101 agccagtttg catctgtcag gatcaatttc ccattatgcc agtcatatta attactagtc
     8161 aattagttga tttttatttt tgacatatac atgtgaa
//
