LOCUS       Exported                6660 bp ds-DNA     circular SYN 22-AUG-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Addgene207876_p0538_GW1_roGFP2_0GG3_mApple
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6660)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Aug 22, 2023 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6660
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             189..868
                     /codon_start=1
                     /note="roGFP2"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLL
                     EFVT"
     misc_feature    384..392
                     /note="Chromophore 65-66-67"
     misc_structure  630..632
                     /note="C147"
     misc_structure  801..803
                     /note="C204"
     CDS             876..1574
                     /codon_start=1
                     /gene="mOrange (variant)"
                     /label=mOrange (variant)
                     /note="mApple"
                     /translation="GEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEAFQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNFEDGG
                     IIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALKSEIK
                     KRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERAEGRH
                     STGGMDELYK"
     polyA_signal    1700..1834
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1858..1874)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1882..1898
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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(1906..1936)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    1951..1972
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2260..2848)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3019..3879)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4533..4549
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5098..5477
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5478..5681
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 aagcttcccg gccacagcaa tcgatccatg acacaaccag tgacaagcag tcccccagtg
       61 aaaccgcatc cgcatccgag tccgataccg ataaagattc tgaatcggag tgagtgccgc
      121 gtccgagagc gttccctgtc cacgtccacc atcggcggag caggtgtgcc tgaggcctct
      181 agaccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      241 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      301 ccacctacgg caagctgacc ctgaagttca tctccaccac cggcaagctg cccgtgccct
      361 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      421 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      481 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      541 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      601 tggggcacaa gctggagtac aactacaact gccacaacgt ctatatcatg gccgacaagc
      661 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      721 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      781 acaaccacta cctgagcacc tgctccgccc tgagcaaaga ccccaacgag aagcgcgatc
      841 acatggtcct gctggagttc gtgaccgctg caggtggaga ggaaaacaac atggccatca
      901 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
      961 agatcgaggg cgagggcgag ggccgcccct acgaggcctt tcagaccgct aagctgaagg
     1021 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     1081 gctccaaggt ctacattaag cacccagccg acatccccga ctacttcaag ctgtccttcc
     1141 ccgagggctt caggtgggag cgcgtgatga acttcgagga cggcggcatt attcacgtta
     1201 accaggactc ctccctgcag gacggcgtgt tcatctacaa ggtgaagctg cgcggcacca
     1261 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccgagg
     1321 agcggatgta ccccgaggac ggcgccctga agagcgagat caagaagagg ctgaagctga
     1381 aggacggcgg ccactacgcc gccgaggtca agaccaccta caaggccaag aagcccgtgc
     1441 agctgcccgg cgcctacatc gtcgacatca agttggacat cgtgtcccac aacgaggact
     1501 acaccatcgt ggaacagtac gaacgcgccg agggtcgtca ctctaccggc ggcatggacg
     1561 agctgtacaa gtaataaact cgacgaattc attgatcata atcagccata ccacatttgt
     1621 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     1681 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     1741 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     1801 caaactcatc aatgtatctt atcatgtctg gatccgtacc gagctcgcgt aatcatggtc
     1861 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1921 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1981 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2041 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2101 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2161 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2221 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2281 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2341 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2401 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     2461 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2521 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2581 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2641 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2701 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2761 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2821 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2881 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2941 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3001 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3061 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3121 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3181 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3241 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3301 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3361 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3421 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3481 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3541 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3601 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3661 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3721 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3781 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3841 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaataagc
     3901 ggccgcggcc atgccggcca ctagtctcga gttattattg aagcatttat cagggttatt
     3961 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4021 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     4081 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     4141 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     4201 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     4261 actatgcggc atcagagcag attgtactga gagtgcacca tatgaagacg tcgcctcctc
     4321 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
     4381 ttagtcagcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     4441 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     4501 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccatctgcag
     4561 tgaataataa aatgtgtgtt tgtccgaaat acgcgtttga gatttctgtc ccgactaaat
     4621 tcatgtcgcg cgatagtggt gtttatcgcc gatagagatg gcgatattgg aaaaatcgat
     4681 atttgaaaat atggcatatt gaaaatgtcg ccgatgtgag tttctgtgta actgatatcg
     4741 ccatttttcc aaaagttgat ttttgggcat acgcgatatc tggcgatacg cttatatcgt
     4801 ttacggggga tggcgataga cgcctttggt gacttgggcg attctgtgtg tcgcaaatat
     4861 cgcagtttcg atataggtga cagacgatat gaggctatat cgccgataga ggcgacatca
     4921 agctggcaca tggccaatgc atatcgatct atacattgaa tcaatattgg ccattagcca
     4981 tattattcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
     5041 atccatatca taatatgtac atttatattg gctcatgtcc aacattaccg ccatgttgac
     5101 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     5161 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     5221 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     5281 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     5341 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     5401 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     5461 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     5521 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     5581 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     5641 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     5701 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
     5761 gcctccgcgg ccgggaacgg tgcattggaa cgcggattcc ccgtgccaag agtgacgtaa
     5821 gtaccgccta tagagtctat aggcccaccc ccttggcttc ttatgcatgc tatactgttt
     5881 ttggcttggg gtctatacac ccccgcttcc tcatgttata ggtgatggta tagcttagcc
     5941 tataggtgtg ggttattgac cattattgac cactccccta ttggtgacga tactttccat
     6001 tactaatcca taacatggct ctttgccaca actctcttta ttggctatat gccaatacac
     6061 tgtccttcag agactgacac ggactctgta tttttacagg atggggtctc atttattatt
     6121 tacaaattca catatacaac accaccgtcc ccagtgcccg cagtttttat taaacataac
     6181 gtgggatctc cacgcgaatc tcgggtacgt gttccggaca tgggctcttc tccggtagcg
     6241 gcggagcttc tacatccgag ccctgctccc atgcctccag cgactcatgg tcgctcggca
     6301 gctccttgct cctaacagtg gaggccagac ttaggcacag cacgatgccc accaccacca
     6361 gtgtgccgca caaggccgtg gcggtagggt atgtgtctga aaatgagctc ggggagcggg
     6421 cttgcaccgc tgacgcattt ggaagactta aggcagcggc agaagaagat gcaggcagct
     6481 gagttgttgt gttctgataa gagtcagagg taactcccgt tgcggtgctg ttaacggtgg
     6541 agggcagtgt agtctgagca gtactcgttg ctgccgcgcg cgccaccaga cataatagct
     6601 gacagactaa cagactgttc ctttccatgg gtcttttctg cagtcaccgt ccttgacacg
//
