LOCUS       sequence-324334-        6825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retro and Lentiviral transduction of primary human germinal center B
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6825)
  AUTHORS   Caeser R, Di Re M, Krupka JA, Gao J, Lara-Chica M, Dias JML, Cooke 
            SL, Fenner R, Usheva Z, Runge HFP, Beer PA, Eldaly H, Pak HK, Park 
            CS, Vassiliou GS, Huntly BJP, Mupo A, Bashford-Rogers RJM, Hodson DJ
  TITLE     Genetic modification of primary human B cells to model high-grade 
            lymphoma.
  JOURNAL   Nat Commun. 2019 Oct 4;10(1):4543. doi: 10.1038/s41467-019-12494-x.
  PUBMED    31586074
REFERENCE   2  (bases 1 to 6825)
  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: "Nat 
            Commun."; date: "2019-10-4"; volume: "10(1):4543. doi"; pages: " 
            10.1038/s41467-019-12494-x"
COMMENT     Sequence Label: phCMV-GALV-MTR
FEATURES             Location/Qualifiers
     source          1..6825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..465
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        466..669
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     620..640
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     666..690
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          781..1353
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(3519..3538)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3584..3639
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3638..3657)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3997..4013)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3997..4013)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4010..4032)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4021..4037
                     /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(4045..4051,4052..4069,4070..4075))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4090..4111
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4228..4245)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4399..4987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4479..4498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5158..5949,5950..6018))
                     /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     5781..5800
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6019..6123)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6149..6604)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6286..6307)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6498..6517
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6731..6753
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6745..6762
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6746..6762
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6772..6791
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6772..6790
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 agagagcttg gcccattgca tacgttgtat ccatatcata atatgtacat ttatattggc
       61 tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca
      121 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      181 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      241 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      301 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      361 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      421 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      481 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      541 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      601 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      661 agcagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac
      721 ctccatagaa gacaccggga ccgatccagc ctccggtcga ccgatcctga gaacttcagg
      781 gtgagtttgg ggacccttga ttgttctttc tttttcgcta ttgtaaaatt catgttatat
      841 ggagggggca aagttttcag ggtgttgttt agaatgggaa gatgtccctt gtatcaccat
      901 ggaccctcat gataattttg tttctttcac tttctactct gttgacaacc attgtctcct
      961 cttattttct tttcattttc tgtaactttt tcgttaaact ttagcttgca tttgtaacga
     1021 atttttaaat tcacttttgt ttatttgtca gattgtaagt actttctcta atcacttttt
     1081 tttcaaggca atcagggtat attatattgt acttcagcac agttttagag aacaattgtt
     1141 ataattaaat gataaggtag aatatttctg catataaatt ctggctggcg tggaaatatt
     1201 cttattggta gaaacaacta caccctggtc atcatcctgc ctttctcttt atggttacaa
     1261 tgatatacac tgtttgagat gaggataaaa tactctgagt ccaaaccggg cccctctgct
     1321 aaccatgttc atgccttctt ctctttccta cagctcctgg gcaacgtgct ggttgttgtg
     1381 ctgtctcatc attttggcaa agaattccgc gggcggccgc tctagaggat ccgctagact
     1441 gacatggtat tgctgcctgg gtccatgctt ctcacctcaa acctgcacca ccttcggcac
     1501 cagatgagtc ctgggagctg gaaaagactg atcatcctct taagctgcgt attcggcggc
     1561 ggcgggacga gtctgcaaaa taagaacccc caccagccca tgaccctcac ttggcaggta
     1621 ctgtcccaaa ctggagacgt tgtctgggat acaaaggcag tccagccccc ttggacttgg
     1681 tggcccacac ttaaacctga tgtatgtgcc ttggcggcta gtcttgagtc ctgggatatc
     1741 ccgggaaccg atgtctcgtc ctctaaacga gtcagacctc cggactcaga ctatactgcc
     1801 gcttataagc aaatcacctg gggagccata gggtgcagct accctcgggc taggactaga
     1861 atggcaagct ctaccttcta cgtatgtccc cgggatggcc ggaccctttc agaagctaga
     1921 aggtgcgggg ggctagaatc cctatactgt aaagaatggg attgtgagac cacggggacc
     1981 ggttattggc tatctaaatc ctcaaaagac ctcataactg taaaatggga ccaaaatagc
     2041 gaatggactc aaaaatttca acagtgtcac cagaccggct ggtgtaaccc ccttaaaata
     2101 gatttcacag acaaaggaaa attatccaag gactggataa cgggaaaaac ctggggatta
     2161 agattctatg tgtctggaca tccaggcgta cagttcacca ttcgcttaaa aatcaccaac
     2221 atgccagctg tggcagtagg tcctgacctc gtccttgtgg aacaaggacc tcctagaacg
     2281 tccctcgctc tcccacctcc tcttccccca agggaagcgc caccgccatc tctccccgac
     2341 tctaactcca cagccctggc gactagtgca caaactccca cggtgagaaa aacaattgtt
     2401 accctaaaca ctccgcctcc caccacaggc gacagacttt ttgatcttgt gcagggggcc
     2461 ttcctaacct taaatgctac caacccaggg gccactgagt cttgctggct ttgtttggcc
     2521 atgggccccc cttattatga agcaatagcc tcatcaggag aggtcgccta ctccaccgac
     2581 cttgaccggt gccgctgggg gacccaagga aagctcaccc tcactgaggt ctcaggacac
     2641 gggttgtgca taggaaaggt gccctttacc catcagcatc tctgcaatca gaccctatcc
     2701 atcaattcct ccggagacca tcagtatctg ctcccctcca accatagctg gtgggcttgc
     2761 agcactggcc tcaccccttg cctctccacc tcagttttta atcagactag agatttctgt
     2821 atccaggtcc agctgattcc tcgcatctat tactatcctg aagaagtttt gttacaggcc
     2881 tatgacaatt ctcaccccag gactaaaaga gaggctgtct cacttaccct agctgtttta
     2941 ctggggttgg gaatcacggc gggaataggt actggttcaa ctgccttaat taaaggacct
     3001 atagacctcc agcaaggcct gacaagcctc cagatcgcca tagatgctga cctccgggcc
     3061 ctccaagact cagtcagcaa gttagaggac tcactgactt ccctgtccga ggtagtgctc
     3121 caaaatagga gaggccttga cttgctgttt ctaaaagaag gtggcctctg tgcggcccta
     3181 aaggaagagt gctgttttta catagaccac tcaggtgcag tacgggactc catgaaaaaa
     3241 ctcaaagaaa aactggataa aagacagtta gagcgccaga aaagccaaaa ctggtatgaa
     3301 ggatggttca ataactcccc ttggtttacc accttgatat ctaccattat gggacccctc
     3361 attgtactcc tattgatttt gctcttcgga ccctgcattc ttaatcgatt agtccaattt
     3421 gttaaagaca ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg
     3481 aagcctatag agtacgagcc atagctcgag ggcccagatc tttttccctc tgccaaaaat
     3541 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     3601 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     3661 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     3721 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     3781 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     3841 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     3901 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     3961 agatccctcg acggatcggc cgcaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     4021 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4081 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4141 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4201 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4261 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4321 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4381 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4441 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4501 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4561 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4621 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4681 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4741 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4801 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4861 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4921 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4981 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5041 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5101 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5161 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5221 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5281 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5341 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5401 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5461 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5521 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5581 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5641 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5701 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5761 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5821 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5881 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5941 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6001 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6061 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6121 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     6181 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     6241 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     6301 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     6361 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     6421 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     6481 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     6541 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     6601 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     6661 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     6721 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac
     6781 tcactatagg gcgaattgga gctccaccgc ggtggcggcc gctct
//
