LOCUS       sequence-137792-        6134 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 6134)
  AUTHORS   Zhou BP, Hu MC, Miller SA, Yu Z, Xia W, Lin SY, Hung MC
  TITLE     HER-2/neu blocks tumor necrosis factor-induced apoptosis via the 
            Akt/NF-kappaB pathway.
  JOURNAL   J Biol Chem. 2000 Mar 17. 275(11):8027-31.
  PUBMED    10713122
REFERENCE   2  (bases 1 to 6134)
  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 Mar 17. 275(11):8027-31."
COMMENT     Sequence Label: HA-Akt DN (K179M)
FEATURES             Location/Qualifiers
     source          1..6134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..140
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     132..149
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     133..149
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        310..689
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        690..893
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     844..864
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             965..991
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2666..2685)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3113..3133)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3118..3447
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3298..3433
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3485..3504)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3486..3504)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3518..3534)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3518..3534)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3531..3553)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3542..3558
                     /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(3566..3572,3573..3590,3591..3596))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3611..3632
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3749..3766)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3920..4508)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4000..4019)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4679..5470,5471..5539))
                     /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     5302..5321
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5540..5644)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5671..6126
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5758..5777)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5968..5989
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
       61 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
      121 agtcacgacg ttgtaaaacg acggccagtg ccaagctgat ctatacattg aatcaatatt
      181 ggcaattagc catattagtc attggttata tagcataaat caatattggc tattggccat
      241 tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac
      301 cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      361 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      421 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      481 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      541 cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat
      601 ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca
      661 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac accaatgggc
      721 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      781 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaataac cccgccccgt
      841 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      901 tgaaccgtca gaattaattc agatctggta ccacgcgtat cgataagctt cgatcgtcga
      961 catgtaccca tacgatgttc cagattacgc ttctagaatg aacgacgtag ccattgtgaa
     1021 ggagggctgg ctgcacaaac gaggggaata tattaaaacc tggcggccac gctacttcct
     1081 cctcaagaac gatggcacct ttattggcta caaggaacgg cctcaggatg tggatcagcg
     1141 agagtcccca ctcaacaact tctcagtggc acaatgccag ctgatgaaga cagagcggcc
     1201 aaggcccaac acctttatca tccgctgcct gcagtggacc acagtcattg agcgcacctt
     1261 ccatgtggaa acgcctgagg agcgggaaga atgggccacc gccattcaga ctgtggccga
     1321 tggactcaag aggcaggaag aagagacgat ggacttccga tcaggctcac ccagtgacaa
     1381 ctcaggggct gaagagatgg aggtgtccct ggccaagccc aagcaccgtg tgaccatgaa
     1441 cgagtttgag tacctgaaac tactgggcaa gggcaccttt gggaaagtga ttctggtgaa
     1501 agagaaggcc acaggccgct actatgccat gatgatcctc aagaaggagg tcatcgtcgc
     1561 caaggatgag gttgcccaca cgcttactga gaaccgtgtc ctgcagaact ctaggcatcc
     1621 cttccttacg gccctcaagt actcattcca gacccacgac cgcctctgct ttgtcatgga
     1681 gtatgccaac gggggcgagc tcttcttcca cctgtctcga gagcgcgtgt tctccgagga
     1741 ccgggcccgc ttctatggtg cggagattgt gtctgccctg gactacttgc actccgagaa
     1801 gaacgtggtg taccgggacc tgaagctgga gaacctcatg ctggacaagg acgggcacat
     1861 caagataacg gacttcgggc tgtgcaagga ggggatcaag gatggtgcca ctatgaagac
     1921 attctgcgga acgccggagt acctggcccc tgaggtgctg gaggacaacg actacggccg
     1981 tgcagtggac tggtgggggc tgggcgtggt catgtatgag atgatgtgtg gccgcctgcc
     2041 cttctacaac caggaccacg agaagctgtt cgagctgatc ctcatggagg agatccgctt
     2101 cccgcgcaca ctcggccctg aggccaagtc cctgctctcc gggctgctca agaaggaccc
     2161 tacacagagg ctcggtgggg gctctgagga tgccaaggag atcatgcagc accggttctt
     2221 tgccaacatc gtgtggcagg atgtgtatga gaagaagctg agcccacctt tcaagcccca
     2281 ggtcacctct gagactgaca ccaggtattt cgatgaggag ttcacagctc agatgatcac
     2341 catcacgccg cctgatcaag atgacagcat ggagtgtgtg gacagtgagc ggaggccgca
     2401 cttcccccag ttctcctact cagccagtgg cacagcctga ggcggatccc cgggcgagct
     2461 cgaattccgt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     2521 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     2581 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     2641 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
     2701 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
     2761 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
     2821 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
     2881 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
     2941 cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca taggctacct
     3001 ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt
     3061 gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct gtggaatgtg
     3121 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3181 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3241 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3301 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3361 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3421 ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt
     3481 aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa
     3541 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3601 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3661 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3721 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3781 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3841 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3901 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3961 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4021 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4081 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4141 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4201 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4261 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4321 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4381 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4441 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4501 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4561 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4621 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4681 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4741 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4801 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4861 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4921 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4981 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5041 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5101 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5161 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5221 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5281 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5341 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5401 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5461 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5521 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5581 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5641 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     5701 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5761 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5821 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5881 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5941 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     6001 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     6061 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     6121 caatttccat tcgc
//
