LOCUS       Exported                7859 bp ds-DNA     circular SYN 24-JUL-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7859)
  AUTHORS   Nathan Shaner
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 25, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7859
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    4..9
                     /label=BglII
     misc_feature    22..27
                     /label=Kozak
     CDS             28..705
                     /codon_start=1
                     /label=mNG d10c
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFT"
     CDS             706..711
                     /codon_start=1
                     /label=GF linker
                     /translation="GF"
     CDS             712..1170
                     /codon_start=1
                     /label=d5n_SSLuc_I13T
                     /translation="EDFVGDWEQTAAYNLDQVLEQGGVSSVLQTLAVSVTPIQRIVRSG
                     ENGLKIDIHVIIPYEGLSADQMAHIEEVFKVVYPVDDHHFKVIMEYGTLVIDGVTPNML
                     NYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLITPEGSMLFRVTIN"
     CDS             1171..1182
                     /codon_start=1
                     /label=SASS linker
                     /translation="SASS"
     CDS             1183..1248
                     /codon_start=1
                     /label=RS20 (from GCaMP6)
                     /translation="DSSRRKWNKTGHAVRAIGRLSS"
     CDS             1249..1293
                     /codon_start=1
                     /label=(GGS)x5 Linker
                     /translation="GGSGGSGGSGGSGGS"
     CDS             1294..1743
                     /codon_start=1
                     /label=GCaMP6F_N294I
                     /translation="NLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRSLGQNP
                     TEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGIGYISA
                     AELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             1744..1752
                     /codon_start=1
                     /label=SPA Linker
                     /translation="SPA"
     CDS             1753..1788
                     /codon_start=1
                     /label=LLEEISN peptide
                     /translation="VTGYRLLEEISN"
     terminator      1900..2127
                     /label=bGH PA terminator
     rep_origin      2190..2496
                     /label=f1 origin
     promoter        2628..2896
                     /label=SV40 promoter
     rep_origin      2795..2872
                     /label=SV40 origin
     terminator      3715..3834
                     /label=SV40 PA terminator
     promoter        complement(3931..3960)
                     /label=lac promoter
     rep_origin      complement(4269..4885)
                     /direction=LEFT
                     /label=pBR322 origin
     CDS             complement(5040..5900)
                     /label=Ampicillin CDS
     promoter        complement(5942..5970)
                     /label=AmpR promoter
     misc_feature    6070..6403
                     /label=CMV enhancer
     promoter        6410..7770
                     /label=Actin promoter
ORIGIN
        1 ctgagatctg catgagctag cgccaccatg gtgagcaagg gcgaggagga taacatggcc
       61 tctctcccag cgacacatga gttacacatc tttggctcca tcaacggtgt ggactttgac
      121 atggtgggtc agggcaccgg caatccaaat gatggttatg aggagttaaa cctgaagtcc
      181 accaagggtg acctccagtt ctccccctgg attctggtcc ctcatatcgg gtatggcttc
      241 catcagtacc tgccctaccc tgacgggatg tcgcctttcc aggccgccat ggtagatggc
      301 tccggatacc aagtccatcg cacaatgcag tttgaagatg gtgcctccct tactgttaac
      361 taccgctaca cctacgaggg aagccacatc aaaggagagg cccaggtgaa ggggactggt
      421 ttccctgctg acggtcctgt gatgaccaac tcgctgaccg ctgcggactg gtgcaggtcg
      481 aagaagactt accccaacga caaaaccatc atcagtacct ttaagtggag ttacaccact
      541 ggaaatggca agcgctaccg gagcactgcg cggaccacct acacctttgc caagccaatg
      601 gcggctaact atctgaagaa ccagccgatg tacgtgttcc gtaagacgga gctcaagcac
      661 tccaagaccg agctcaactt caaggagtgg caaaaggcct ttaccggatt tgaggacttc
      721 gttggtgact gggagcagac cgctgcttac aacctggacc aggttctgga acagggtggt
      781 gtgtcttctg tgctccagac gctggctgtt tctgtgactc cgatccagcg tatcgttcgt
      841 tctggtgaaa acggtctgaa aatcgacatc cacgttatca tcccgtacga aggtctgtct
      901 gctgatcaga tggctcatat cgaagaggtc ttcaaagttg tttacccggt tgacgaccac
      961 cacttcaaag tcatcatgga gtacggtacc ctggttatcg acggtgttac cccgaacatg
     1021 ctcaactact tcggtcgtcc gtacgagggt atcgctgttt tcgacggtaa gaagatcacc
     1081 gttaccggta ccctgtggaa cggtaacaaa atcatcgacg aacgtctgat cacccctgaa
     1141 ggttctatgc tgttccgtgt taccatcaac tctgcttctt ctgactcatc acgtcgtaag
     1201 tggaataaga caggtcacgc agtcagagct ataggtcggc tgagctcagg tggatccggt
     1261 ggtagtggtg gatccggagg ttctggtgga tccaacctgc cggaccaact gactgaagag
     1321 cagatcgcag aatttaaaga ggagttctcc ctatttgaca aggacgggga tgggacaata
     1381 acaaccaagg agctggggac ggtgatgcgg tctctggggc agaaccccac agaagcagag
     1441 ctgcaggaca tgatcaatga agtagatgcc gacggtgacg gcacaatcga cttccctgag
     1501 ttcctgacaa tgatggcaag aaaaatgaaa tacagggaca cggaagaaga aattagagaa
     1561 gcgttcggtg tgtttgataa ggatggcatt ggctacatca gtgcagcaga gcttcgccac
     1621 gtgatgacaa accttggaga gaagttaaca gatgaagagg ttgatgaaat gatcagggaa
     1681 gcagacatcg atggggatgg tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg
     1741 aagtctcctg ctgttaccgg ttaccgtctg ctcgaagaaa tctctaacta aggtaccgag
     1801 ctcggatcca ctagtccagt gtggtggaat tctgcagata tccagcacag tggcggccgc
     1861 tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     1921 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     1981 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2041 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2101 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     2161 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2221 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2281 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2341 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2401 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2461 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2521 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2581 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2641 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2701 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2761 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2821 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2881 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2941 caaaaagctc ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     3001 ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     3061 gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     3121 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     3181 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     3241 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     3301 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     3361 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     3421 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     3481 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     3541 cccggtgcct gattcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     3601 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     3661 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     3721 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3781 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3841 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     3901 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3961 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4021 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4081 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4141 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4201 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4261 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4321 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4381 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4441 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4501 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4561 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4621 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4681 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4741 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4801 caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4861 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4921 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4981 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5041 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5101 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5161 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5221 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5281 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5341 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5401 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5461 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5521 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5581 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5641 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5701 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5761 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5821 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5881 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5941 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6001 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcggggtc gacattgatt
     6061 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     6121 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     6181 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     6241 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     6301 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     6361 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     6421 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
     6481 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
     6541 gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg
     6601 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
     6661 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc
     6721 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
     6781 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
     6841 cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc
     6901 cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg
     6961 gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
     7021 gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg
     7081 tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga
     7141 gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt
     7201 gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc
     7261 gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc
     7321 ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg
     7381 gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg
     7441 tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc
     7501 ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg
     7561 ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc
     7621 cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga
     7681 gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
     7741 gttattgtgc tgtctcatca ttttggcaaa gaattcctcg aggaattcac tcctcaggtg
     7801 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaatacca
//
