LOCUS       Exported                8728 bp ds-DNA     circular SYN 06-SEP-2018
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8728)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Dec 14, 2020 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8728
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             905..1621
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1:  905 ..  907 / #05fd14
                        2:  908 ..  910 / #05fd14 / 1a
                        3:  911 .. 1621 / #05fd14"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     protein_bind    1643..1667
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
                     /note="color: #31849b; direction: RIGHT"
     primer_bind     1682..1703
                     /label=Insert.FOR
                     /note="color: black; sequence: atgtcggcgccgtcggaggagg"
     primer_bind     complement(4233..4249)
                     /label=Insert.REV
                     /note="color: black; sequence: ctaatcacagtaaatgc"
     protein_bind    complement(4263..4287)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
                     /note="color: #31849b; direction: LEFT"
     polyA_signal    4325..4549
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      4595..5023
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        5037..5366
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      5217..5352
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             5433..6227
                     /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    6401..6522
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(6571..6587)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    6595..6611
                     /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(6619..6649)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 6619 .. 6625 / #ffffff / -10
                        2: 6626 .. 6643 / #ffffff
                        3: 6644 .. 6649 / #ffffff / -35"
     protein_bind    6664..6685
                     /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(6973..7561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(7732..8592)
                     /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: 7732 .. 8523 / #ccffcc
                        2: 8524 .. 8592 / #ccffcc / signal sequence
                     Cleavage site after base 8523"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8593..8697)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctaga
      901 caccatggcc agcaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
      961 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgctac
     1021 atacggaaag cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc
     1081 aacacttgtc actactttct cttatggtgt tcaatgcttt tcccgttatc cggatcatat
     1141 gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg aacgcactat
     1201 atctttcaaa gatgacggga actacaagac gcgtgctgaa gtcaagtttg aaggtgatac
     1261 ccttgttaat cgtatcgagt taaaaggtat tgattttaaa gaagatggaa acattctcgg
     1321 acacaaactc gagtacaact ataactcaca caatgtatac atcacggcag acaaacaaaa
     1381 gaatggaatc aaagctaact tcaaaattcg ccacaacatt gaagatggat ccgttcaact
     1441 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
     1501 ccattacctg tcgacacaat ctgccctttc gaaagatccc aacgaaaagc gtgaccacat
     1561 ggtccttctt gagtttgtaa ctgctgctgg gattacacat ggcatggatg agctctacaa
     1621 aagcggttgg cctccggatc aaacaagttt gtacaaaaaa gcaggctccg aattcgccct
     1681 tatgtcggcg ccgtcggagg aggaggagta cgcgcggctg gtgatggagg cgcagccgga
     1741 gtggctgcgc gccgaggtga agcggctgtc ccacgagctg gccgagacca cgcgtgagaa
     1801 gatccaggcg gccgagtacg ggctggcggt gctcgaggag aagcaccagc tcaagctgca
     1861 gttcgaggag ctcgaggtgg actatgaggc tatccgcagc gagatggagc agctcaagga
     1921 ggcctttgga caagcacaca caaaccacaa gaaggtggct gctgacggag agagccggga
     1981 ggagagcctg atccaggagt cggcctccaa ggagcagtac tacgtgcgga aggtgctaga
     2041 gctgcagacg gagctgaagc agttgcgcaa tgtcctcacc aacacgcagt cggagaatga
     2101 gcgcctggcc tctgtggccc aggagctgaa ggagatcaac cagaatgtgg agatccagcg
     2161 tggccgcctg cgggatgaca tcaaggagta caaattccgg gaagctcgtc tgctgcagga
     2221 ctactcggaa ctggaggagg agaacatcag cctgcagaag caagtgtctg tgctcagaca
     2281 gaaccaggtg gagtttgagg gcctcaagca tgagatcaag cgtctggagg aggagaccga
     2341 gtacctcaac agccagctgg aggatgccat ccgcctcaag gagatctcag agcggcagct
     2401 ggaggaggcg ctggagaccc tgaagacgga gcgcgaacag aagaacagcc tgcgcaagga
     2461 gctgtcacac tacatgagca tcaatgactc cttctacacc agccacctgc atgtctcgct
     2521 ggatggcctc aagttcagtg acgatgctgc cgagcccaac aacgatgccg aggccctggt
     2581 caatggcttt gagcacggcg gcctggccaa gctgccactg gacaacaaga cctccacgcc
     2641 caagaaggag ggcctcgcac cgccctcccc cagcctcgtc tccgacctac tcagtgagct
     2701 caacatctct gagatccaga agctgaagca gcagctgatg cagatggagc gggaaaaggc
     2761 gggcctgctg gcaacgctgc aggacacaca gaagcagctg gagcacacgc ggggctccct
     2821 gtcagaacag caggagaagg tgacccgcct cacagagaat ctgagtgccc tgcggcgcct
     2881 gcaggccagc aaggagcggc agacagccct ggacaacgag aaggaccgtg acagccatga
     2941 ggatggggac tactacgagg tggacatcaa cgggcctgag atcttggcct gcaagtacca
     3001 tgtggctgtg gctgaggctg gcgagctccg cgagcagctc aaggcactgc gcagcacgca
     3061 cgaggctcgt gaggcccagc acgccgagga gaagggccgc tatgaggctg agggccaggc
     3121 actcacggag aaggtctccc tgctagagaa ggccagccgc caggaccgcg agctgctggc
     3181 ccggctggag aaggagctaa agaaggtgag cgacgtcgcc ggcgagacac agggcagcct
     3241 gagtgtggcc caggatgagc tggtgacctt cagtgaggag ctggccaatc tctaccacca
     3301 cgtgtgcatg tgcaacaatg agacacccaa ccgtgtcatg ctggactact accgcgaggg
     3361 ccagggcggg gccggccgca ccagtcccgg gggccgcacc agccccgagg cgcgtggccg
     3421 gcgctcaccc atcctcctac ccaaggggct gctggctcct gaggcgggcc gagcagatgg
     3481 tgggacgggg gacagcagcc cctcgcctgg ctcctcactg ccatcacccc tgagtgaccc
     3541 acgccgggag cccatgaaca tctacaacct gatcgctatc atccgtgacc agatcaagca
     3601 cctgcaggca gccgtggacc gcaccacgga gctgtcacgc cagcgcattg cctctcagga
     3661 gctgggcccc gccgtggaca aggacaagga agcgcttatg gaggagatcc tcaagctgaa
     3721 gtcgctgctc agcaccaagc gggagcagat caccacgctg cgcactgtgc tcaaggccaa
     3781 caagcagacg gccgaggtgg cccttgccaa cctgaagagc aagtatgaga atgagaaggc
     3841 catggttacc gagaccatga tgaagctgcg caatgagctc aaggccctca aggaggacgc
     3901 agccaccttc tcctcgctgc gtgctatgtt tgccaccagg tgtgacgagt acattacaca
     3961 gctggatgag atgcagcggc agctggcggc tgctgaggac gagaagaaga cgctgaactc
     4021 gctgctgcgc atggccatcc agcagaagct ggcgctgacc cagcggctgg agctgctcga
     4081 gctggaccat gagcagaccc ggcgtggccg tgccaaagcc gccccgaaga ccaagccagc
     4141 cacaccgagc gtaagtcaca cctgtgcctg tgccagcgac agggccgagg gcaccgggct
     4201 ggccaaccag gtgttctgca gcgagaagca cagcatttac tgtgattaga agggcgaatt
     4261 cgacccagct ttcttgtaca aagtggtgat aattaattaa gataaacccg ctgatcagcc
     4321 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     4381 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4441 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4501 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     4561 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4621 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4681 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4741 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4801 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4861 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4921 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4981 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     5041 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5101 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5161 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5221 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5281 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     5341 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     5401 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5461 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5521 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5581 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5641 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5701 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5761 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5821 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5881 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5941 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6001 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6061 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6121 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6181 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6241 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6301 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6361 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     6421 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6481 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     6541 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6601 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6661 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6721 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6781 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6841 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6901 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6961 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7021 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7081 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7141 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7201 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7261 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7321 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7381 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7441 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7501 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7561 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7621 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7681 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7741 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7801 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7861 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7921 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7981 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8041 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8101 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8161 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8221 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8281 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8341 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8401 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8461 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8521 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8581 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8641 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8701 atttccccga aaagtgccac ctgacgtc
//
