LOCUS       sequence-335109-        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(SS32,36AA)
FEATURES             Location/Qualifiers
     source          1..5909
                     /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    1760..2382
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1959..1978)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2406..2426)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2411..2740
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2591..2726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          2779..2875
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     primer_bind     complement(2951..2970)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2952..2970)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2984..3000)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2984..3000)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2997..3019)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3008..3024
                     /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(3032..3038,3039..3056,3057..3062))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3077..3098
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3215..3232)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3386..3974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3466..3485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4145..4936,4937..5005))
                     /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     4768..4787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5006..5110)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5137..5592
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5224..5243)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5434..5455
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5718..5740
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5732..5749
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5733..5749
                     /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 gataagctta ccatgttcca ggcggccgag cgcccccagg agtgggccat ggagggcccc
      841 cgcgacgggc tgaagaagga gcggctactg gacgaccgcc acgacgccgg cctggacgca
      901 atgaaagacg aggagtacga gcagatggtc aaggagctgc aggagatccg cctcgagccg
      961 caggaggtgc cgcgcggctc ggagccctgg aagcagcagc tcaccgagga cggggactcg
     1021 ttcctgcact tggccatcat ccatgaagaa aaggcactga ccatggaagt gatccgccag
     1081 gtgaagggag acctggcctt cctcaacttc cagaacaacc tgcagcagac tccactccac
     1141 ttggctgtga tcaccaacca gccagaaatt gctgaggcac ttctgggagc tggctgtgat
     1201 cctgagctcc gagactttcg aggaaatacc cccctacacc ttgcctgtga gcagggctgc
     1261 ctggccagcg tgggagtcct gactcagtcc tgcaccaccc cgcacctcca ctccatcctg
     1321 aaggctacca actacaatgg ccacacgtgt ctacacttag cctctatcca tggctacctg
     1381 ggcatcgtgg agcttttggt gtccttgggt gctgatgtca atgctcagga gccctgtaat
     1441 ggccggactg cccttcacct cgcagtggac ctgcaaaatc ctgacctggt gtcactcctg
     1501 ttgaagtgtg gggctgatgt caacagagtt acctaccagg gctattctcc ctaccagctc
     1561 acctggggcc gcccaagcac ccggatacag cagcagctgg gccagctgac actagaaaac
     1621 cttcagatgc tgccagagag tgaggatgag gagagctatg acacagagtc agagttcacg
     1681 gagttcacag aggacgagct gccctatgat gactgtgtgt ttggaggcca gcgtctgacg
     1741 ttatgatcta gaggatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     1801 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     1861 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     1921 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     1981 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     2041 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     2101 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     2161 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     2221 tcccttccct gtccttctga ttttaaaata actataccag caggaggacg tccagacaca
     2281 gcataggcta cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt
     2341 cctctcatgc gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcttg
     2401 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2461 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2521 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2581 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2641 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     2701 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcctcg aggaactgaa
     2761 aaaccagaaa gttaactggt aagtttagtc tttttgtctt ttatttcagg tcccggatcc
     2821 ggtggtggtg caaatcaaag aactgctcct cagtggatgt tgcctttact tctaggcctg
     2881 tacggaagtg ttacttctgc tctaaaagct gctgcaggtc gactctaggg ccgcgtaccg
     2941 agctcgaatt ccctatagtg agtcgtatta aattcgtaat catgtcatag ctgtttcctg
     3001 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3061 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3121 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3181 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3241 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3301 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3361 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3421 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3481 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3541 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3601 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3661 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3721 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3781 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3841 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3901 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3961 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4021 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4081 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4141 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4201 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4261 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4321 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4381 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4441 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4501 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4561 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4621 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4681 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4741 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4801 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4861 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4921 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4981 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5041 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5101 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa
     5161 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     5221 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5281 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5341 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5401 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5461 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     5521 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5581 cgcttacaat ttccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     5641 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     5701 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctgat
     5761 ctatacattg aatcaatatt ggcaattagc catattagtc attggttata tagcataaat
     5821 caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat
     5881 tggctcatgt ccaatatgac cgccatgtt
//
