LOCUS       sequence-449174-        6447 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 6447)
  AUTHORS   Toney LM, Cattoretti G, Graf JA, Merghoub T, Pandolfi PP, 
            Dalla-Favera R, Ye BH, Dent AL
  TITLE     BCL-6 regulates chemokine gene transcription in macrophages.
  JOURNAL   Nat Immunol. 2000 Sep;1(3):214-20.
  PUBMED    10973278
REFERENCE   2  (bases 1 to 6447)
  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 
            Immunol."; date: "2000-09"; volume: "1(3)"; pages: "214-20"
COMMENT     Sequence Label: pMCP-MU-Luc (MCP-1 mut promoter in pGL3-basic)
FEATURES             Location/Qualifiers
     source          1..6447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    587..635
                     /note="synthetic polyadenylation signal"
     misc_feature    649..740
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     689..708
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     enhancer        768..1071
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1226..1246
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2458..2467
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2464..4116
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(2530..2548)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4157..4278
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4168..4187)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4222..4241
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4526..4543)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4697..5285)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4777..4796)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5456..6247,6248..6316))
                     /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     6079..6098
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6317..6421)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat
      541 gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata aaatatcttt
      601 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct
      661 ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc
      721 aggtgccaga acatttctct atcgataggt accgagctct tacgcgtcgt tacataactt
      781 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      841 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      901 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      961 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     1021 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     1081 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     1141 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     1201 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggac
     1261 gcgtgctagc ccgggtccgc tcagccactg gcacgcatgt gagagcgcca ctctttttat
     1321 caagagtctg ctgaaatgaa ttctaagggc tttcagattt tatcgcttcg atcacactgt
     1381 ttgtgaagaa atctaaacct ggaaagctga attaagccag acattccagt tggctcactc
     1441 atggtccctg gaaatcacga aggaaacagg gcagagagct accaggagta gtaaacgcta
     1501 acacacaggc ttctaattca tcatggctta accaaaaggc atgtttctcc atggcccgtt
     1561 agctgtctgt tagtttgaag gtaatgtgat gatagccagg ctggagctac ctgggtcctg
     1621 ttccagggac tcttagggca gttacctaca tgacccttct ggacctcagc tgcctgatct
     1681 gagaatgggg agtaacagca tctacttacc agagctgggg aaattagaga ctgcactgtg
     1741 aagggttcca accactcctg gcccagagta agcactaggt gagttttata tagaaatttc
     1801 ttctgcacca tgagctcatg tatatggctt tccaggtctg gacttttaca actcagacag
     1861 ttcatatcaa tgtcacaaag cttcttagtg ctgcaaaata tctggtaatc accaagtgga
     1921 gagaatgctg tattcaacaa ggcctgataa ccaaggactc agccgactta ctgggggtcc
     1981 tttcccagct gcaaggttcc ttgagccagg ggcaagctag gctgtgtttc caagcatctt
     2041 cttcctaagg actttaggtt ccttggccac ttccttttat ttcagtgaaa gcagagccac
     2101 tccattcaca cttgtggtca cagtagtaca attactgcca attcttccct ctttcccccc
     2161 ccccccccta ctccctgcgc agcttcattt gctcccagga gtggctagaa aaataccaaa
     2221 ttccaaccca cagtttctct cttccacttc ctttaaacac ccgagggctc tgcacttact
     2281 cagcggattc aacttccact ttccatcact tatccagggt gatgctactc cttggcacca
     2341 agcaccctgc ctgactccac ccccctggct tacaataaaa ggctgcctca gagcagccag
     2401 aagtgcactc gagatctgcg atctaagtaa gcttggcatt ccggtactgt tggtaaagcc
     2461 accatggaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc gctggaagat
     2521 ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca
     2581 attgctttta cagatgcaca tatcgaggtg gacatcactt acgctgagta cttcgaaatg
     2641 tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc
     2701 gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga
     2761 gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgggc
     2821 atttcgcagc ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg
     2881 caaaaaaagc tcccaatcat ccaaaaaatt attatcatgg attctaaaac ggattaccag
     2941 ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac
     3001 gattttgtgc cagagtcctt cgatagggac aagacaattg cactgatcat gaactcctct
     3061 ggatctactg gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg cgtgagattc
     3121 tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt
     3181 gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga
     3241 tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttctgaggag ccttcaggat
     3301 tacaagattc aaagtgcgct gctggtgcca accctattct ccttcttcgc caaaagcact
     3361 ctgattgaca aatacgattt atctaattta cacgaaattg cttctggtgg cgctcccctc
     3421 tctaaggaag tcggggaagc ggttgccaag aggttccatc tgccaggtat caggcaagga
     3481 tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg
     3541 ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg
     3601 aaaacgctgg gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc
     3661 ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat
     3721 tctggagaca tagcttactg ggacgaagac gaacacttct tcatcgttga ccgcctgaag
     3781 tctctgatta agtacaaagg ctatcaggtg gctcccgctg aattggaatc catcttgctc
     3841 caacacccca acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa
     3901 cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg
     3961 gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg
     4021 gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc
     4081 ataaaggcca agaagggcgg aaagatcgcc gtgtaattct agagtcgggg cggccggccg
     4141 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4201 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4261 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     4321 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     4381 gatccgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     4441 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     4501 ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4561 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4621 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4681 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4741 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4801 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4861 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4921 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4981 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5041 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5101 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5161 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5221 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5281 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5341 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5401 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5461 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5521 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5581 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5641 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5701 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5761 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5821 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5881 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5941 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6001 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6061 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6121 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6181 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6241 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6301 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6361 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6421 gcacatttcc ccgaaaagtg ccacctg
//
