LOCUS       sequence-352645-        6935 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 6935)
  AUTHORS   Dickerson SK, Market E, Besmer E, Papavasiliou FN
  TITLE     AID mediates hypermutation by deaminating single stranded DNA.
  JOURNAL   J Exp Med. 2003 May 19. 197(10):1291-6.
  PUBMED    12756266
REFERENCE   2  (bases 1 to 6935)
  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: "J Exp Med. 
            2003 May 19. 197(10):1291-6."
COMMENT     Sequence Label: pMSCVgfp::AID
FEATURES             Location/Qualifiers
     source          1..6935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             66..582
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    646..987
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1054..1470
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1400..1422
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1438..1454
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1489..2085
                     /codon_start=1
                     /gene="AICDA"
                     /product="human activation-induced cytidine deaminase 
                     (Dickerson et al., 2003)"
                     /label=AID
                     /translation="MDSLLMNRRKFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLD
                     FGYLRNKNGCHVELLFLRYISDWDLDPGRCYRVTWFTSWSPCYDCARHVADFLRGNPNL
                     SLRIFTARLYFCEDRKAEPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHERTFKAWE
                     GLHENSVRLSRQLRRILLPLYEVDDLRDAFRTLGL"
     promoter        2099..2598
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2134..2157)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2531..2550
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      2616..2625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2622..2624,2625..2627,2628..3341)
                     /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(2622..2624,2625..2627,2628..3338)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2667..2688)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2928..2947)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3275..3296
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3395..3909
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4071..4087
                     /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(4095..4101,4102..4119,4120..4125))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4140..4161
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4278..4295)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4449..5037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4529..4548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5208..5999,6000..6068))
                     /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     5831..5850
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6069..6173)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6241..6259
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6297..6319)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6419..6438
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6632..6654
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6782..6801
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aagcctatag agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag
       61 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
      121 aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga
      181 gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
      241 gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca
      301 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
      361 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
      421 acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtatt
      481 cccaataaag cctcttgctg tttgcatccg aatcgtggac tcgctgatcc ttgggagggt
      541 ctcctcagat tgattgactg cccacctcgg gggtctttca tttggaggtt ccaccgagat
      601 ttggagaccc ctgcccaggg accaccgacc cccccgccgg gaggtaagct ggccagcggt
      661 cgtttcgtgt ctgtctctgt ctttgtgcgt gtttgtgccg gcatctaatg tttgcgcctg
      721 cgtctgtact agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag
      781 ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt
      841 ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg
      901 gtaggagacg agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg
      961 aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt
     1021 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
     1081 ttgaccttag gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc
     1141 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     1201 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     1261 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     1321 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct
     1381 ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt
     1441 tatccagccc tcactccttc tctaggcgcc ggaattagat ctctcgagat ggacagcctc
     1501 ttgatgaacc ggaggaagtt tctttaccaa ttcaaaaatg tccgctgggc taagggtcgg
     1561 cgtgagacct acctgtgcta cgtagtgaag aggcgtgaca gtgctacatc cttttcactg
     1621 gactttggtt atcttcgcaa taagaacggc tgccacgtgg aattgctctt cctccgctac
     1681 atctcggact gggacctaga ccctggccgc tgctaccgcg tcacctggtt cacctcctgg
     1741 agcccctgct acgactgtgc ccgacatgtg gccgactttc tgcgagggaa ccccaacctc
     1801 agtctgagga tcttcaccgc gcgcctctac ttctgtgagg accgcaaggc tgagcccgag
     1861 gggctgcggc ggctgcaccg cgccggggtg caaatagcca tcatgacctt caaagattat
     1921 ttttactgct ggaatacttt tgtagaaaac catgaaagaa ctttcaaagc ctgggaaggg
     1981 ctgcatgaaa attcagttcg tctctccaga cagcttcggc gcatcctttt gcccctgtat
     2041 gaggttgatg acttacgaga cgcatttcgt actttgggac tttgatgaga attctaccgg
     2101 gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc
     2161 acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc
     2221 caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg
     2281 aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt
     2341 ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg
     2401 ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt
     2461 gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc
     2521 ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc
     2581 tcatctccgg gcctttcgac ctgcagccca agcttgccac catggtgagc aagggcgagg
     2641 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     2701 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     2761 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     2821 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2881 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2941 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     3001 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     3061 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     3121 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     3181 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     3241 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     3301 ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccat cgataaaata
     3361 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     3421 caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag
     3481 aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg
     3541 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc
     3601 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     3661 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     3721 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga
     3781 tagactgcgt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact
     3841 tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     3901 gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga
     3961 atattaagta atcctgactc aattagccac tgttttgaat ccacatactc caatactcct
     4021 gaaatagttc attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg
     4081 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4141 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4201 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4261 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4321 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4381 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4441 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4501 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4561 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4621 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4681 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4741 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4801 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4861 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4921 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4981 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5041 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5101 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5161 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5221 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5281 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5341 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5401 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5461 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5521 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5581 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5641 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5701 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5761 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5821 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5881 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5941 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6001 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6061 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6121 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6181 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     6241 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     6301 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     6361 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     6421 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     6481 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     6541 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     6601 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg
     6661 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     6721 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     6781 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     6841 cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc
     6901 aggctctagt tttgactcaa caatatcacc agctg
//
