LOCUS       sequence-421233-        4970 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 4970)
  AUTHORS   Xiao G, Sun SC
  TITLE     Negative regulation of the nuclear factor kappa B-inducing kinase by
            a cis-acting domain.
  JOURNAL   J Biol Chem. 2000 Jul 14. 275(28):21081-5.
  PUBMED    10887201
REFERENCE   2  (bases 1 to 4970)
  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 Biol 
            Chem. 2000 Jul 14. 275(28):21081-5."
COMMENT     Sequence Label: pCMV4-HA
FEATURES             Location/Qualifiers
     source          1..4970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             682..771
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     polyA_signal    821..1443
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1020..1039)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1467..1487)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1472..1801
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1652..1787
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          1840..1936
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     primer_bind     complement(2012..2031)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2013..2031)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2045..2061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2045..2061)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2058..2080)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2069..2085
                     /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(2093..2099,2100..2117,2118..2123))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2138..2159
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2276..2293)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2447..3035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2527..2546)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3206..3997,3998..4066))
                     /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     3829..3848
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4067..4171)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4198..4653
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4285..4304)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4495..4516
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4779..4801
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4793..4810
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4794..4810
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgttt ttatttttaa ttttctttca aatacttcca tcgaattcag atctggtacc
      661 acgcgtacca tggatatctt ttacccatac gatgttcctg actatgcggg ctatccctat
      721 gacgtcccgg actatgcagg atcctatcca tatgacgttc cagattacgc tgctcagatc
      781 gataagcttg catgcctgca ggtcgactct agaggatccc gggtggcatc cctgtgaccc
      841 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
      901 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
      961 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     1021 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     1081 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     1141 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     1201 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     1261 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca
     1321 gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt
     1381 gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga
     1441 attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     1501 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     1561 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     1621 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     1681 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     1741 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     1801 aaagctcctc gaggaactga aaaaccagaa agttaactgg taagtttagt ctttttgtct
     1861 tttatttcag gtcccggatc cggtggtggt gcaaatcaaa gaactgctcc tcagtggatg
     1921 ttgcctttac ttctaggcct gtacggaagt gttacttctg ctctaaaagc tgctgcaggt
     1981 cgactctagg gccgcgtacc gagctcgaat tccctatagt gagtcgtatt aaattcgtaa
     2041 tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2101 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2161 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2221 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2281 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2341 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2401 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2461 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2521 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2581 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2641 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2701 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2761 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2821 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2881 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2941 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3001 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3061 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3121 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3181 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3241 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3301 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3361 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3421 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3481 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3541 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3601 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3661 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3721 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3781 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3841 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3901 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3961 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4021 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4081 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4141 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4201 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4261 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4321 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4381 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4441 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4501 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4561 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4621 cgaattttaa caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg
     4681 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     4741 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     4801 gacggccagt gccaagctga tctatacatt gaatcaatat tggcaattag ccatattagt
     4861 cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata
     4921 tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt
//
