LOCUS       sequence-480525-        7471 bp    DNA     circular SYN 25-JUL-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7471)
  AUTHORS   Wang ZY, Sato H, Kusam S, Sehra S, Toney LM, Dent AL
  TITLE     Regulation of IL-10 gene expression in Th2 cells by Jun proteins.
  JOURNAL   J Immunol. 2005 Feb 15;174(4):2098-105.
  PUBMED    15699140
REFERENCE   2  (bases 1 to 7471)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 25, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J 
            Immunol."; date: "2005-02-15"; volume: "174(4)"; pages: "2098-105"
COMMENT     Sequence Label: pMIEG3-JunB
FEATURES             Location/Qualifiers
     source          1..7471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             60..576
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    640..981
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1048..1464
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1394..1416
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1432..1448
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(2540..3114,3115..3117,3118..3126)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2707..2724)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2934..2953
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3127..3129,3130..3132,3133..3843)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3127..3129,3130..3132,3133..3843)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3172..3193)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3433..3452)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3780..3801
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3925..4439
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4601..4617
                     /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(4625..4631,4632..4649,4650..4655))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4670..4691
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4808..4825)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4979..5567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5059..5078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5738..6529,6530..6598))
                     /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     6361..6380
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6599..6703)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6771..6789
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6827..6849)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6949..6968
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7162..7184
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7312..7331
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atagagtacg agccatagat aaaataaaag attttattta gtctccagaa aaagggggga
       61 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
      121 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
      181 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
      241 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
      301 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      361 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      421 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      481 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      541 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      601 acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      661 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      721 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      781 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      841 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      901 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      961 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
     1021 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1081 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1141 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1201 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1261 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1321 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1381 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1441 gccctcactc cttctctagg cgccggaatt ctgtgcacga aaatggaaca gcctttctat
     1501 cacgacgact cttacgcagc ggcgggatac ggtcggagcc ctggcagcct gtctctacac
     1561 gactacaaac tcctgaaacc caccttggcg ctcaacctgg cggatcccta tcggggtctc
     1621 aagggtcctg gggcgcgggg tccaggcccg gagggcagtg gggcaggcag ctacttttcg
     1681 ggtcagggat cagacacagg cgcatctctg aagctagcct ccacggaact ggagcgcttg
     1741 atcgtcccca acagcaacgg cgtgatcacg acgacgccca cgcctccggg acagtacttt
     1801 tacccccgtg ggggtggcag cggtggaggt acagggggcg gcgtcaccga ggagcaggag
     1861 ggctttgcgg acggttttgt caaagccctg gacgacctgc acaagatgaa ccacgtgacg
     1921 ccccccaacg tgtccctggg cgccagcggg ggtccccagg ccggcccagg gggcgtctat
     1981 gctggtccgg agccgcctcc cgtctacacc aacctcagca gttactctcc agcctctgca
     2041 ccctctggag gctccgggac cgccgtcggg actgggagct catacccgac ggccaccatc
     2101 agctacctcc cacatgcacc accctttgcg ggcggccacc cggcacagct gggtttgagt
     2161 cgtggcgctt ccgcctttaa agaggaaccg cagaccgtac cggaggcacg cagccgcgac
     2221 gccacgccgc ctgtgtcccc catcaacatg gaagaccagg agcgcatcaa agtggagcga
     2281 aagcggctgc ggaacaggct ggcggccacc aagtgccgga agcggaagct ggagcgcatc
     2341 gcgcgcctgg aggacaaggt gaagacactc aaggctgaga acgcggggct gtcgagtgct
     2401 gccggtctcc taagggagca agtggcgcag ctcaagcaga aggtcatgac ccatgtcagc
     2461 aacggctgcc agttgctgct aggggtcaag ggacacgcct tctgagaatt ccaaggcctt
     2521 ggcggccgct cgagatccgc ccctctccct cccccccccc taacgttact ggccgaagcc
     2581 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     2641 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     2701 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     2761 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     2821 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     2881 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     2941 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     3001 ctgatctggg gcctcggtac acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     3061 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
     3121 acaaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     3181 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     3241 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     3301 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     3361 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     3421 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     3481 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3541 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     3601 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     3661 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3721 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     3781 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3841 aagtccggac tcagatccgt cgacctgcag ccaagcttat cgataaaata aaagatttta
     3901 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     3961 taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat
     4021 caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag
     4081 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca
     4141 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     4201 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     4261 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
     4321 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     4381 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg
     4441 ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta
     4501 atcctgactc aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc
     4561 attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg ctcacaattc
     4621 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4681 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4741 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4801 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4861 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4921 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4981 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5041 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5101 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5161 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5221 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5281 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5341 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5401 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5461 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5521 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5581 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5641 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5701 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5761 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5821 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5881 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5941 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6001 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6061 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6121 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6181 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6241 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6301 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6361 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6421 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6481 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6541 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6601 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6661 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6721 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6781 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     6841 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     6901 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     6961 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     7021 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     7081 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     7141 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg caaggaatgg
     7201 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     7261 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     7321 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     7381 ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt
     7441 tttgactcaa caatatcacc agctgaagcc t
//
