LOCUS       Untitled_36        7798 bp DNA     circular SYN 23-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7798)
  AUTHORS   Addgene
  TITLE     Direct Submission
  JOURNAL   Exported Apr 23, 2025 from SnapGene 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        192..1480
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             join(1501..1503,1504..1506,1507..2217)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1501..1503,1504..1506,1507..2217)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2227..2280
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(2290..2292,2293..2313,2314..3585)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="APKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAAE
                     LTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKL
                     IPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIW
                     EITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVI
                     IQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQ
                     LLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSD
                     KRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLK
                     GSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     misc_feature    3613..4201
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4084..4095)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4234..4458
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4679..5008)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4693..4828
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(5254..5842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6013..6804,6805..6873))
                     /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(6874..6978)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        7766..7784
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 ttctagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
       61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      121 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      181 tgctctagta acattatggc cttaggtcac ttcatctcca tggggttctt cttctgattt
      241 tctagaaaat gagatggggg tgcagagagc ttcctcagtg acctgcccag ggtcacatca
      301 gaaatgtcag agctagaact tgaactcaga ttactaatct taaattccat gccttggggg
      361 catgcaagta cgatatacag aaggagtgaa ctcattaggg cagatgacca atgagtttag
      421 gaaagaagag tccagggcag ggtacatcta caccacccgc ccagccctgg gtgagtccag
      481 ccacgttcac ctcattatag ttgcctctct ccagtcctac cttgacggga agcacaagca
      541 gaaactggga caggagcccc aggagaccaa atcttcatgg tccctctggg aggatgggtg
      601 gggagagctg tggcagaggc ctcaggaggg gccctgctgc tcagtggtga cagatagggg
      661 tgagaaagca gacagagtca ttccgtcagc attctgggtc tgtttggtac ttcttctcac
      721 gctaaggtgg cggtgtgata tgcacaatgg ctaaaaagca gggagagctg gaaagaaaca
      781 aggacagaga cagaggccaa gtcaaccaga ccaattccca gaggaagcaa agaaaccatt
      841 acagagacta caagggggaa gggaaggaga gatgaattag cttcccctgt aaaccttaga
      901 acccagctgt tgccagggca acggggcaat acctgtctct tcagaggaga tgaagttgcc
      961 agggtaacta catcctgtct ttctcaagga ccatcccaga atgtggcacc cactagccgt
     1021 taccatagca actgcctctt tgccccactt aatcccatcc cgtctgttaa aagggcccta
     1081 tagttggagg tgggggaggt aggaagagcg atgatcactt gtggactaag tttgttcgca
     1141 tccccttctc caaccccctc agtacatcac cctgggggaa cagggtccac ttgctcctgg
     1201 gcccacacag tcctgcagta ttgtgtatat aaggccaggg caaagaggag caggttttaa
     1261 agtgaaaggc aggcaggtgt tggggaggca gttaccgggg caacgggaac agggcgtttc
     1321 ggaggtggtt gccatgggga cctggatgct gacgaaggct cgcgaggctg tgagcagcca
     1381 cagtgccctg ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt ttgcactcag
     1441 gagcacgggc aggcgagtgg cccctagttc tgggggcagc gggggatccg tcgacccacc
     1501 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1561 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1621 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1681 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     1741 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1801 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1861 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1921 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     1981 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     2041 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2101 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     2161 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc
     2221 agtggagagg gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa tcctggccca
     2281 gcggccgcag ctcctaagaa gaagaggaag gtgatgagcc agttcgacat cctgtgcaag
     2341 acccccccca aggtgctggt gcggcagttc gtggagagat tcgagaggcc cagcggcgag
     2401 aagatcgcca gctgtgccgc cgagctgacc tacctgtgct ggatgatcac ccacaacggc
     2461 accgccatca agagggccac cttcatgagc tacaacacca tcatcagcaa cagcctgagc
     2521 ttcgacatcg tgaacaagag cctgcagttc aagtacaaga cccagaaggc caccatcctg
     2581 gaggccagcc tgaagaagct gatccccgcc tgggagttca ccatcatccc ttacaacggc
     2641 cagaagcacc agagcgacat caccgacatc gtgtccagcc tgcagctgca gttcgagagc
     2701 agcgaggagg ccgacaaggg caacagccac agcaagaaga tgctgaaggc cctgctgtcc
     2761 gagggcgaga gcatctggga gatcaccgag aagatcctga acagcttcga gtacaccagc
     2821 aggttcacca agaccaagac cctgtaccag ttcctgttcc tggccacatt catcaactgc
     2881 ggcaggttca gcgacatcaa gaacgtggac cccaagagct tcaagctggt gcagaacaag
     2941 tacctgggcg tgatcattca gtgcctggtg accgagacca agacaagcgt gtccaggcac
     3001 atctactttt tcagcgccag aggcaggatc gaccccctgg tgtacctgga cgagttcctg
     3061 aggaacagcg agcccgtgct gaagagagtg aacaggaccg gcaacagcag cagcaacaag
     3121 caggagtacc agctgctgaa ggacaacctg gtgcgcagct acaacaaggc cctgaagaag
     3181 aacgccccct accccatctt cgctatcaag aacggcccta agagccacat cggcaggcac
     3241 ctgatgacca gctttctgag catgaagggc ctgaccgagc tgacaaacgt ggtgggcaac
     3301 tggagcgaca agagggcctc cgccgtggcc aggaccacct acacccacca gatcaccgcc
     3361 atccccgacc actacttcgc cctggtgtcc aggtactacg cctacgaccc catcagcaag
     3421 gagatgatcg ccctgaagga cgagaccaac cccatcgagg agtggcagca catcgagcag
     3481 ctgaagggca gcgccgaggg cagcatcaga taccccgcct ggaacggcat catcagccag
     3541 gaggtgctgg actacctgag cagctacatc aacaggcgga tctgagaatt caagctttaa
     3601 cagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3661 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3721 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3781 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3841 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3901 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3961 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     4021 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4081 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4141 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4201 catcgatacc gtcgactcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     4261 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     4321 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     4381 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     4441 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg actagagcat
     4501 ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg
     4561 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     4621 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagagctttt
     4681 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg
     4741 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg
     4801 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
     4861 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     4921 acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     4981 gggactttcc acaccctaac tgacacacat tccacagctg cattaatgaa tcggccaacg
     5041 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5101 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5161 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5221 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5281 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5341 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5401 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5461 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5521 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5581 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5641 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5701 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5761 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5821 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5881 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5941 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6001 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6061 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6121 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6181 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6241 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6301 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6361 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6421 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6481 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6541 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6601 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6661 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6721 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6781 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6841 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6901 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6961 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     7021 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     7081 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     7141 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     7201 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattc
     7261 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     7321 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     7381 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     7441 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     7501 cggtgatgcc ggccacgatg cgtccggcgt agaggatctg gctagcgatg accctgctga
     7561 ttggttcgct gaccatttcc gggtgcggga cggcgttacc agaaactcag aaggttcgtc
     7621 caaccaaacc gactctgacg gcagtttacg agagagatga tagggtctgc ttcagtaagc
     7681 cagatgctac acaattaggc ttgtacatat tgtcgttaga acgcggctac aattaataca
     7741 taaccttatg tatcatacac atacgattta ggtgacacta tagaatacac ggaattaa
//
