LOCUS       Exported                8547 bp ds-DNA     circular SYN 14-JUL-2018
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8547)
  AUTHORS   Hochgeschwender
  TITLE     Direct Submission
  JOURNAL   Exported Jul 22, 2018 from SnapGene 4.1.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8547
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        38..1761
                     /label=CAG promoter
                     /note="color: #ffffff"
     misc_feature    1777..1786
                     /label=Kozak sequence
                     /note="color: #a6acb3"
     CDS             1783..1833
                     /codon_start=1
                     /label=Gluc Signal Sequence
                     /note="color: #ff0000"
                     /translation="MGVKVLFALICIAVAEA"
     CDS             1834..2343
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nanoluc
                     /note="human codon-optimized"
                     /note="color: #00ccff"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     CDS             2344..2388
                     /codon_start=1
                     /label=15 aa Linker
                     /note="color: #993300"
                     /translation="GDPLVQCGGIAGSAT"
     CDS             2389..3288
                     /codon_start=1
                     /label=VChR1
                     /note="color: #993366"
                     /translation="MDYPVARSLIVRYPTDLGNGTVCMPRGQCYCEGWLRSRGTSIEKT
                     IAITLQWVVFALSVACLGWYAYQAWRATCGWEEVYVALIEMMKSIIEAFHEFDSPATLW
                     LSSGNGVVWMRYGEWLLTCPVLLIHLSNLTGLKDDYSKRTMGLLVSDVGCIVWGATSAM
                     CTGWTKILFFLISLSYGMYTYFHAAKVYIEAFHTVPKGICRELVRVMAWTFFVAWGMFP
                     VLFLLGTEGFGHISPYGSAIGHSILDLIAKNMWGVLGNYLRVKIHEHILLYGDIRKKQK
                     ITIAGQEMEVETLVAEEED"
     CDS             3298..4014
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 2 segments:
                        1: 3298 .. 3300 / #e1ff2f / 1a
                        2: 3301 .. 4014 / #e1ff2f"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             4044..4085
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /note="color: #cc99b2"
                     /translation="GKPIPNPLLGLDST"
     CDS             4095..4112
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     polyA_signal    4141..4365
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      4411..4839
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4853..5183
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      5034..5169
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             5250..6044
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    6220..6341
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(6390..6406)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    6414..6430
                     /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)."
                     /note="color: #31849b"
     promoter        complement(6438..6468)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 6438 .. 6444 / #ffffff / -10
                        2: 6445 .. 6462 / #ffffff
                        3: 6463 .. 6468 / #ffffff / -35"
     protein_bind    6483..6504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(6792..7380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(7551..8411)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 7551 .. 8342 / #ccffcc
                        2: 8343 .. 8411 / #ccffcc / signal sequence
                     Cleavage site after base 8342"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8412..8516)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctggc gcgccggtac catgcattag ttattaatag taatcaatta
       61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      361 cttggcagta catctacgta ttagtcatcg ctattaccat gggtcgaggt gagccccacg
      421 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
      481 ttttaattat tttgtgcagc gatgggggcg gggggggggg gggcgcgcgc caggcggggc
      541 ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag
      601 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
      661 gcgaagcgcg cggcgggcgg gagtcgctgc gttgccttcg ccccgtgccc cgctccgcgc
      721 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
      781 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc tcgtttcttt
      841 tctgtggctg cgtgaaagcc ttaaagggct ccgggagggc cctttgtgcg ggggggagcg
      901 gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc ccgcgctgcc
      961 cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgcg tgtgcgcgag
     1021 gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg ctgcgagggg aacaaaggct
     1081 gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcggcg gtcgggctgt
     1141 aacccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg
     1201 ctccgtgcgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg
     1261 gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc
     1321 cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat
     1381 cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg gcggagccga aatctgggag
     1441 gcgccgccgc accccctcta gcgggcgcgg gcgaagcggt gcggcgccgg caggaaggaa
     1501 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct
     1561 cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
     1621 ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc gttcttcttt
     1681 ttctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaag
     1741 ctagcgctac cggactcaga tctcgagctc aagcttgcca ccatgggagt caaagttctg
     1801 tttgccctga tctgcatcgc tgtggccgag gccgtcttca cactcgaaga tttcgttggg
     1861 gactggcgac agacagccgg ctacaacctg gaccaagtcc ttgaacaggg aggtgtgtcc
     1921 agtttgtttc agaatctcgg ggtgtccgta actccgatcc aaaggattgt cctgagcggt
     1981 gaaaatgggc tgaagatcga catccatgtc atcatcccgt atgaaggtct gagcggcgac
     2041 caaatgggcc agatcgaaaa aatttttaag gtggtgtacc ctgtggatga tcatcacttt
     2101 aaggtgatcc tgcactatgg cacactggta atcgacgggg ttacgccgaa catgatcgac
     2161 tatttcggac ggccgtatga aggcatcgcc gtgttcgacg gcaaaaagat cactgtaaca
     2221 gggaccctgt ggaacggcaa caaaattatc gacgagcgcc tgatcaaccc cgacggctcc
     2281 ctgctgttcc gagtaaccat caacggagtg accggctggc ggctgtgcga acgcattctg
     2341 gcgggcgacc cactagtcca gtgtggtgga attgccggat ccgccaccat ggattaccct
     2401 gttgctagaa gcctcatagt tcgctaccca accgacctcg gaaacggcac cgtctgcatg
     2461 ccaagaggac agtgttactg tgaaggttgg cttcggagtc gcggcacttc cattgaaaag
     2521 acaatagcaa ttactcttca gtgggtagtc tttgctttgt cagtggcttg cctggggtgg
     2581 tatgcgtatc aagcgtggcg agctacctgc ggatgggagg aggtttacgt agccttgata
     2641 gaaatgatga aaagcatcat cgaggccttc cacgagttcg acagccctgc aacactgtgg
     2701 ctgtcttcag ggaacggcgt agtttggatg cggtatggcg aatggctcct cacctgcccg
     2761 gtccttctga tccatctgag caacctcaca ggcctgaagg acgattatag caaaaggact
     2821 atgggcctgt tggtttctga tgtgggatgc atcgtgtggg gcgcaaccag cgccatgtgt
     2881 acggggtgga cgaagatcct gttcttcctc atctcattga gctatggtat gtatacctat
     2941 tttcatgctg ctaaagttta tatcgaagca ttccacacag ttccaaaagg gatttgtcga
     3001 gaactggtcc gagtgatggc ctggacattc tttgtggctt ggggaatgtt tccagtcctg
     3061 tttctgctgg gcacggaagg attcggtcat atcagccctt atggatctgc cattgggcac
     3121 tccatcctcg acctgattgc aaagaacatg tggggtgtgc tggggaatta cctgcgcgtc
     3181 aaaatccacg agcacatcct gttgtatggc gacatcagaa agaagcagaa aattacgatc
     3241 gccggccaag agatggaggt tgagacactg gtggctgaag aggaggacgc ggccgccgtg
     3301 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     3361 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     3421 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     3481 accaccttcg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     3541 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     3601 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     3661 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     3721 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     3781 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     3841 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3901 agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3961 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaacatatg
     4021 ctctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt
     4081 ctacgcgtac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     4141 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     4201 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     4261 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     4321 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     4381 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     4441 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4501 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4561 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4621 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4681 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4741 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt
     4801 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     4861 gttagggtgt ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc
     4921 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4981 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5041 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5101 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     5161 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     5221 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5281 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5341 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5401 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5461 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5521 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5581 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5641 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5701 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5761 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5821 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5881 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5941 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6001 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6061 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     6121 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     6181 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     6241 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6301 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     6361 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6421 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6481 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6541 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6601 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6661 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6721 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6781 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6841 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6901 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6961 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     7021 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7081 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7141 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7201 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7261 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7321 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7381 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7441 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7501 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7561 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7621 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7681 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7741 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7801 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7861 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7921 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7981 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8041 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8101 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8161 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8221 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8281 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8341 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8401 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8461 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8521 tttccccgaa aagtgccacc tgacgtc
//
