LOCUS       Exported                7319 bp ds-DNA     circular SYN 15-AUG-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7319)
  AUTHORS   NakahataY
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Aug 21, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7319
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        19..398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        400..678
                     /label=chicken beta-actin promoter
     intron          679..1699
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     regulatory      1769..1778
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1781..1807
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1811..1837
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1841..1867
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1880..4321
                     /codon_start=1
                     /product="https://www.ncbi.nlm.nih.gov/nucleotide/NM_001432
                     039.1"
                     /label=Evi5l_NM_001432039.1
                     /translation="SLSTMASPTLSPDSSSQEALSAPTCSPTSDSENLSPDELELLAKL
                     EEQNRLLEADSKSMRSMNGSRRNSGSSLVSSSSASSNLSHLEEDTWILWGRIANEWEEW
                     RRRKEKLLKELIRKGIPHHFRAIVWQLLCSATDMPVKNQYSELLKMSSPCEKLIRRDIA
                     RTYPEHEFFKGQDSLGQEVLFNVMKAYSLVDREVGYCQGSAFIVGLLLMQMPEEEAFCV
                     FVRLMQEYRLRELFKPSMAELGLCIYQFEYMLQEQLPDLNTHFRSQSFHTSMYASSWFL
                     TLFLTTFPLPVATRVFDIFMYEGLEIVFRVGLALLQVNQTELMQLDMEGMSQYFQRVIP
                     HQFDSCPDKLVLKAYQVKYNPKKMKRLEKEYAAMKSKEMEEQIEIKRLRTENRLLKQRI
                     ETLEKESAALADRLIQGQVTRAQEAEENYVIKRELAVVRQQCSSTAEDLQKAQSTIRQL
                     QEQQENPRLTEDFVSHLETELEQSRLRETETLGALREMQDKVLDMEKRNSSLPDENNVA
                     RLQEELKALKVREGEAVASARELKLQLQELSDTWQAHVSRGGRWKESPRKLVLGELQDE
                     LMTMRLREAQALADGREWRQRVVELETQDNIHRNLLNRVEAERAALQEKLQYLAAQNKG
                     LQTQLSESRRKQAEAECKSKEEVMAVRLREADSMAAVAEMRQRIAELEIQREEGRIQGQ
                     LNHSDSSQYIRELKDQIEELKTEVRLLKGPPTFEDPLAFDGLSLTRHLDEDSLPSSDEE
                     LLGVGVGVGVGAALQDPLYPLSPRDARFFRSLDRPAKDSEGSSDSDADELATPYSNQVL
                     DN"
     polyA_signal    4416..4471
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4832..4848)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4856..4872
                     /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(4880..4910)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4925..4946
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5005..5200
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5051..5186
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5206..5340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5579..6167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6338..7198)
                     /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"
     promoter        complement(7199..7303)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg
       61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
      421 gcttcactct ccccatctcc ccccccctcc ccacccccaa ttttgtattt atttattttt
      481 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg
      541 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
      601 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      661 cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc
      721 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg
      781 cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct
      841 tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc
      901 ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg
      961 cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc
     1021 gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa
     1081 ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg
     1141 ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg
     1201 gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg
     1261 ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc
     1321 ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg
     1381 taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct
     1441 gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg
     1501 aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc
     1561 cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg
     1621 ttcggcttct ggcgtgtgac cggcggctct agctagagcc tctgctaacc atgttcatgc
     1681 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt
     1741 tggcaaagaa ttcgctagcg ctaccggtgc caccatggca tacccttatg acgtaccaga
     1801 ttacgcggga tatccctacg atgtccccga ttacgcgcgt tatccatatg acgtcccgga
     1861 ctatgccgaa ttcctcgaga gcctgtccac catggcaagc cccactctga gccccgactc
     1921 ctcatctcag gaggccctgt cagcacccac ctgctcccca acctctgact ccgagaacct
     1981 cagcccagat gagctagaac tactggccaa gcttgaggag cagaaccggc tcctggaagc
     2041 cgactccaag tccatgcgct ccatgaatgg ctcccggagg aacagcggct cctccctggt
     2101 atccagctcc tccgcctcct ccaacctgag ccacctggag gaggacacat ggattctctg
     2161 gggccggatt gccaacgagt gggaggaatg gcggcgcaga aaggaaaagc tactcaagga
     2221 gctgatccga aagggcatcc cacaccactt ccgggccatc gtttggcagc tcctgtgcag
     2281 tgccacagat atgcccgtca aaaaccagta ctccgagctc ctcaagatgt cctcaccatg
     2341 tgagaagctg atccgaaggg acattgcccg cacctatcca gagcatgagt tcttcaaagg
     2401 ccaggacagc ctgggccagg aagtcctctt caatgtcatg aaggcatact ccctggttga
     2461 tcgggaggtt ggatattgcc aaggcagtgc cttcattgtg ggcttactcc tcatgcagat
     2521 gcctgaggaa gaggccttct gtgtgtttgt gcggctgatg caggagtacc ggctacggga
     2581 gctcttcaag cccagcatgg ccgagctggg actctgtatc taccagtttg aatacatgct
     2641 gcaggagcag cttccggacc tgaacaccca cttccgctca cagagcttcc acacatccat
     2701 gtatgcatca tcctggttcc tcacgctttt cctcaccacc tttcccctgc ctgttgctac
     2761 ccgggtcttt gacatcttta tgtatgaggg actggagatt gtgttccgag tcggccttgc
     2821 cctgctgcaa gtgaaccaga cagagctgat gcagctggac atggagggca tgtcccagta
     2881 cttccagaga gtgatccccc accagtttga cagctgccca gacaagctgg tcctcaaggc
     2941 ttaccaggtc aagtacaacc ctaaaaagat gaagaggctg gagaaggagt acgcagctat
     3001 gaagagtaag gagatggagg agcagatcga gatcaaaagg cttcgaacag agaaccggct
     3061 gctcaaacag cggattgaga ccctagagaa ggagagcgcg gccttggctg ataggttaat
     3121 ccagggacaa gtgacacggg cacaggaggc tgaggagaac tatgtcatca agcgggagtt
     3181 ggcagtagtg agacaacaat gtagctcgac tgcagaggac cttcagaaag cacagagtac
     3241 cattcggcag ttgcaagaac agcaggaaaa ccctcgccta acagaggact ttgtttccca
     3301 cctggagaca gaactggagc agtcacggtt acgggagacc gagactctgg gggcccttcg
     3361 agaaatgcag gacaaagttc tggacatgga aaagagaaac agctcactgc cggatgagaa
     3421 caatgtagcc aggctgcagg aagaactcaa ggccctcaag gtgcgggagg gcgaggccgt
     3481 agcctctgcg cgggaactga agctgcagct gcaggagctc tcagacacct ggcaggctca
     3541 tgtttcccgc ggtggccgct ggaaggagtc cccgcggaag ctggtcctgg gagagttaca
     3601 ggatgagctc atgactatgc gtttgcgtga ggcacaggcc ctggcggatg gccgtgagtg
     3661 gcggcagcgc gtggtggaac tggagacgca ggacaacatc caccgcaacc tgctgaaccg
     3721 ggtggaggcg gaacgcgcgg ctctgcagga gaagttgcag tacctggcag cacagaacaa
     3781 gggactccag actcagctca gcgaaagccg ccgcaagcag gccgaggctg agtgcaagag
     3841 taaggaggaa gtgatggctg tgcgccttcg ggaggcagac agcatggctg cagtggcgga
     3901 gatgaggcag cgcatcgcag agctggagat ccagagggag gagggccgca tccagggcca
     3961 gctgaaccat tcggattcat cgcagtacat ccgtgagctg aaggaccaga ttgaggagct
     4021 gaagactgag gttcggttgc tgaaaggccc acccacattc gaggacccgc tggctttcga
     4081 tgggctgagc ctaacgcggc acctggacga ggattcgcta ccgtcttctg atgaggagct
     4141 gcttggtgtg ggtgtgggcg tgggcgtggg cgcagccctg caggacccac tctaccctct
     4201 gtctccccgg gacgcacgct tcttcagaag tctggaccgg ccagccaagg acagcgaagg
     4261 cagctcagac agcgatgcag atgagctagc cacgccttac agcaaccagg tcctggacaa
     4321 ctgagcggcc gcgactctag agtcgacgga tctttttccc tctgccaaaa attatgggga
     4381 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     4441 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     4501 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     4561 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     4621 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     4681 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     4741 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     4801 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4861 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4921 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4981 gaaacctgtc gtgccagcgg atacgcatct caattagtca gcaaccatag tcccgcccct
     5041 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     5101 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     5161 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca
     5221 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5281 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctcgatc
     5341 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5401 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5461 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5521 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5581 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5641 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5701 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5761 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5821 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5881 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5941 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6001 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6061 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6121 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6181 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6241 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6301 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6361 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6421 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6481 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6541 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6601 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6661 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6721 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6781 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6841 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6901 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6961 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7021 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7081 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7141 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7201 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7261 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaa
//
