LOCUS       sequence-217589-        5909 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 5909)
  AUTHORS   Sun S, Elwood J, Greene WC
  TITLE     Both amino- and carboxyl-terminal sequences within I kappa B alpha 
            regulate its inducible degradation.
  JOURNAL   Mol Cell Biol. 1996 Mar . 16(3):1058-65.
  PUBMED    8622650
REFERENCE   2  (bases 1 to 5909)
  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: "Mol Cell 
            Biol. 1996 Mar . 16(3):1058-65."
COMMENT     Sequence Label: pCMV4-3 HA/IkB-alpha
FEATURES             Location/Qualifiers
     source          1..5909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        38..417
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        418..621
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     572..592
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             719..808
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     polyA_signal    1797..2419
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1996..2015)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2443..2463)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2448..2777
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2628..2763
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          2816..2912
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     primer_bind     complement(2988..3007)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2989..3007)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3021..3037)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3021..3037)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3034..3056)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3045..3061
                     /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(3069..3075,3076..3093,3094..3099))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3114..3135
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3252..3269)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3423..4011)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3503..3522)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4182..4973,4974..5042))
                     /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     4805..4824
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5043..5147)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5174..5629
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5261..5280)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5471..5492
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5755..5777
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5769..5786
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5770..5786
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atttatattg gctcatgtcc aatatgaccg ccatgttgac attgattatt gactagttat
       61 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      121 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      181 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      241 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtccg
      301 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      361 ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      421 atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg gggatttcca
      481 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      541 ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg
      601 gaggtctata taagcagagc tcgtttagtg aaccgtcaga attgttttta tttttaattt
      661 tctttcaaat acttccatcg aattcagatc tggtaccacg cgtaccatgg atatctttta
      721 cccatacgat gttcctgact atgcgggcta tccctatgac gtcccggact atgcaggatc
      781 ctatccatat gacgttccag attacgctgc tcagatcgat aagcttacca tgttccaggc
      841 ggccgagcgc ccccaggagt gggccatgga gggcccccgc gacgggctga agaaggagcg
      901 gctactggac gaccgccacg acagcggcct ggactccatg aaagacgagg agtacgagca
      961 gatggtcaag gagctgcagg agatccgcct cgagccgcag gaggtgccgc gcggctcgga
     1021 gccctggaag cagcagctca ccgaggacgg ggactcgttc ctgcacttgg ccatcatcca
     1081 tgaagaaaag gcactgacca tggaagtgat ccgccaggtg aagggagacc tggccttcct
     1141 caacttccag aacaacctgc agcagactcc actccacttg gctgtgatca ccaaccagcc
     1201 agaaattgct gaggcacttc tgggagctgg ctgtgatcct gagctccgag actttcgagg
     1261 aaataccccc ctacaccttg cctgtgagca gggctgcctg gccagcgtgg gagtcctgac
     1321 tcagtcctgc accaccccgc acctccactc catcctgaag gctaccaact acaatggcca
     1381 cacgtgtcta cacttagcct ctatccatgg ctacctgggc atcgtggagc ttttggtgtc
     1441 cttgggtgct gatgtcaatg ctcaggagcc ctgtaatggc cggactgccc ttcacctcgc
     1501 agtggacctg caaaatcctg acctggtgtc actcctgttg aagtgtgggg ctgatgtcaa
     1561 cagagttacc taccagggct attctcccta ccagctcacc tggggccgcc caagcacccg
     1621 gatacagcag cagctgggcc agctgacact agaaaacctt cagatgctgc cagagagtga
     1681 ggatgaggag agctatgaca cagagtcaga gttcacggag ttcacagagg acgagctgcc
     1741 ctatgatgac tgtgtgtttg gaggccagcg tctgacgtta tgatctagag gatcccgggt
     1801 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     1861 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     1921 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     1981 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     2041 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     2101 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     2161 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     2221 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     2281 taaaataact ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc
     2341 aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc
     2401 agtagatgcc tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag
     2461 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2521 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2581 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2641 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2701 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2761 gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aactggtaag
     2821 tttagtcttt ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaagaac
     2881 tgctcctcag tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct
     2941 aaaagctgct gcaggtcgac tctagggccg cgtaccgagc tcgaattccc tatagtgagt
     3001 cgtattaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3061 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3121 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3181 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3241 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3301 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3361 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3421 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3481 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3541 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3601 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3661 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3721 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3781 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3841 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3901 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3961 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4021 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4081 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4141 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4201 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4261 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4321 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4381 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4441 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4501 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4561 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4621 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4681 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4741 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4801 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4861 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4921 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4981 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5041 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5101 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5161 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5221 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5281 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5341 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5401 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5461 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     5521 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     5581 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat
     5641 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     5701 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     5761 cacgacgttg taaaacgacg gccagtgcca agctgatcta tacattgaat caatattggc
     5821 aattagccat attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc
     5881 atacgttgta tctatatcat aatatgtac
//
