LOCUS       Exported                6308 bp ds-DNA     circular SYN 08-NOV-2017
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    NC90 Doc2b GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6308)
  AUTHORS   Nick Courtney
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jun 18, 2019 from SnapGene Viewer 4.3.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6308
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        36..1217
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          266..1208
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     CDS             1251..2486
                     /codon_start=1
                     /label=rDoc2b
                     /translation="MTLRRRGEKATISIQEHMAIDVCPGPIRPIKQISDYFPRFPRGLP
                     PTAAPRASAPPDAPARSPAATAGPRSPSDGARDDDEDVDQLFGAYGASPGPSPGPSPVR
                     PPAKPPEDEPDADGYESDDCTALGTLDFSLLYDQENNALHCTISKAKGLKPMDHNGLAD
                     PYVKLHLLPGASKANKLRTKTLRNTLNPSWNETLTYYGITDEDMIRKTLRISVCDEDKF
                     RHNEFIGETRVPLKKLKPNHTKTFSICLEKQLPVDKAEDKSLEERGRILISLKYSSQKQ
                     GLLVGIVRCAHLAAMDANGYSDPYVKTYLKPDVDKKSKHKTAVKKKTLNPEFNEEFCYE
                     IKHGDLAKKTLEVTVWDYDIGKSNDFIGGVVLGINAKGERLKHWFDCLKNKDKRIERWH
                     TLTNEIPGAVLSD"
     misc_structure  1629..2003
                     /label=C2A ncbi
     misc_feature    1716..1739
                     /label=C2A Loop 1
     misc_feature    1902..1928
                     /label=C2A Loop 3
     misc_structure  2019..2375
                     /label=C2B ncbi
     misc_feature    2136..2159
                     /label=C2B Loop 1
     misc_feature    2319..2345
                     /label=C2B Loop 3
     CDS             2529..3248
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_site      3397..3452
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(3811..3827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3835..3851
                     /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(3859..3889)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3904..3925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3984..4179
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4030..4165
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4185..4319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4558..5146)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5317..6177)
                     /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(6178..6282)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agatcatcgc gtgaggctcc ggtgcccgtc agtgggcaga
       61 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc
      121 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
      181 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
      241 caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct
      301 ctttacgggt tatggccctt gcgtgccttg aattacttcc acgcccctgg ctgcagtacg
      361 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
      421 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
      481 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
      541 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
      601 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
      661 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
      721 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
      781 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
      841 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
      901 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
      961 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
     1021 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
     1081 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
     1141 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
     1201 catttcaggt gtcgtgagga attctgcagt cgacggtacc gcgggcccgg atgaccctcc
     1261 gaagacgtgg agagaaggct acaatcagca tccaagagca tatggcaatc gacgtgtgtc
     1321 ctggacctat ccgacctatc aagcagatct ccgattactt cccacgattc cctcgtggac
     1381 tcccaccaac agctgcacca cgtgcttcag cacctccaga cgctcctgca cgatcgccag
     1441 cagctaccgc aggtcctcgt agcccctccg acggagcacg cgacgacgac gaagatgtgg
     1501 accagctctt cggagcctac ggtgccagcc caggccccag ccccggaccc agccccgtga
     1561 ggccgccggc caagcccccc gaggacgaac cggacgccga cggctacgag tcagacgact
     1621 gcaccgccct gggtacactg gacttcagtc tgctctatga ccaggagaac aacgcactgc
     1681 actgcaccat cagcaaagcc aagggcctga agccgatgga tcacaatgga ctggctgatc
     1741 cctacgtcaa actacacttg ctgcctggag ccagcaaggc aaataagctc agaacaaaaa
     1801 ccctacggaa cactctgaac ccctcctgga acgagaccct cacctattac gggatcacgg
     1861 atgaggacat gatccgaaag accctgagga tctccgtgtg tgacgaggac aaattccgcc
     1921 acaacgagtt catcggagag actcgagtgc ccctgaagaa gctgaaaccc aaccacacca
     1981 agacattcag catctgcctg gagaagcagc tgccggtgga caaggcagag gacaagtccc
     2041 tggaggagcg tggccgcatc ctcatctctc tcaagtacag ctcacagaag cagggcctgc
     2101 tggtgggcat cgttcgctgc gcacacctgg cggccatgga tgctaacggc tactcagacc
     2161 cctacgtgaa aacatatctg aaaccagatg tagacaagaa atccaagcat aagaccgcag
     2221 tcaagaagaa aacactaaac cctgaattca atgaggagtt ctgttacgag atcaagcatg
     2281 gagacctagc caaaaagacc ctggaggtca ctgtctggga ctatgatatt ggaaaatcca
     2341 atgatttcat cggtggggtg gttctgggca tcaatgccaa gggtgagcgc ctgaagcact
     2401 ggtttgactg tctgaagaac aaggacaaga ggattgagcg ttggcacacg ctcaccaatg
     2461 agatcccagg ggctgtactc agcgacggat ccggaagttc ggggtccagc ggttcttcag
     2521 gcagttccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     2581 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     2641 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     2701 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     2761 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     2821 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2881 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2941 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     3001 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     3061 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     3121 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     3181 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     3241 acaagtaagg ccgcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     3301 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     3361 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     3421 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     3481 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc atatgctggc
     3541 tgccatgaac aaaggtggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     3601 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     3661 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     3721 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg aagatccctc
     3781 gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3841 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3901 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3961 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta
     4021 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4081 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4141 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag
     4201 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4261 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc
     4321 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4381 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4441 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4501 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4561 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4621 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4681 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4741 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4801 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4861 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4921 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4981 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5041 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5101 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5161 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5221 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5281 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5341 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5401 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5461 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5521 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5581 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5641 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5701 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5761 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5821 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5881 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5941 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6001 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6061 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6121 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6181 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6241 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6301 gccacctg
//
