LOCUS       Exported                8433 bp ds-DNA     circular SYN 09-APR-2019
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8433)
  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..8433
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     regulatory      670..679
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             679..708
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDL"
     CDS             724..753
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDL"
     protein_bind    772..796
                     /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"
     protein_bind    814..838
                     /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     853..876
                     /label=Insert.FOR
                     /note="color: black; sequence: atggagctggaggacggtgtggtg"
     primer_bind     complement(4792..4818)
                     /label=Insert.REV
                     /note="color: black; sequence: tcactcattgccatacatcacttgcca"
     protein_bind    complement(4832..4856)
                     /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"
     promoter        complement(4935..4953)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    5227..5348
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(5355..5810)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        5837..5941
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        5943..6300
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      6151..6286
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             6335..7129
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    7361..7408
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /note="color: #a6acb3"
     rep_origin      7737..8325
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
      661 gcggtggcgg ccgccatgga gcagaaactc atctctgaag aggatctgag cccgggcgga
      721 tccgagcaga aactcatctc tgaagaggat ctgctgcagg aattcgatat cacaagtttg
      781 tacaaaaaag caggctccga attcgccctt atcacaagtt tgtacaaaaa agcaggctcc
      841 gaattcgccc ttatggagct ggaggacggt gtggtgtatc aggaggagcc cggcggctcc
      901 ggggccgtga tgtcggagcg ggtgtccggc ctggccggct ccatctaccg cgagttcgag
      961 cggcttatcg ggcgctatga cgaggaggtg gtcaaagagc tgatgccgct ggtggtggct
     1021 gtgctggaga acctggactc ggtgttcgcg caggaccagg agcaccaggt ggagctggag
     1081 ctgctgcggg acgacaacga gcagctcatc acccagtacg agcgggagaa ggcgctgcgc
     1141 aagcacgctg aggagaaatt cattgaattt gaagactctc aagaacagga aaaaaaggac
     1201 ttacagaccc gagtggaatc tttagaatct caaacaagac aacttgagct gaaagcgaaa
     1261 aactatgctg accagattag cagacttgaa gaaagagaag cagaactgaa gaaggaatat
     1321 aatgcattac atcaaagaca cactgagatg atccataatt atatggaaca tttagaaaga
     1381 acaaaacttc atcagctctc agggagtgat caactagaat ccacagctca tagtagaatt
     1441 agaaaagaac gccctatatc attaggaatt ttcccattac ctgctggaga tggattgctt
     1501 acacctgatg ctcagaaagg aggagagacc cctggatctg agcaatggaa atttcaggaa
     1561 ttaagtcaac cacgttctca taccagcctg aaggtcagca atagtcctga acctcagaag
     1621 gctgtagaac aggaggatga gctttctgat gttagccaag gcggatctaa agctaccact
     1681 ccagcatcaa cagctaattc agatgtggca acaattccta ctgatactcc cttaaaggaa
     1741 gaaaacgaag gatttgtgaa ggttacagat gcgccaaata aatcagagat aagcaaacac
     1801 attgaagtac aggtagccca ggaaactaga aatgtatcta ctggctctgc tgaaaatgaa
     1861 gaaaagtcag aagttcaagc aatcatcgaa tctactcctg agctggatat ggacaaagat
     1921 ctcagtggat ataaaggttc aagcactccc accaaaggca tagagaacaa agcttttgat
     1981 cgcaatacag aatctctctt tgaagaactg tcttcagctg gctcaggcct aataggagat
     2041 gtggatgaag gagcagattt actaggaatg ggtcgggaag ttgagaatct tatattagaa
     2101 aatacacaac tgttggaaac caaaaatgct ttgaacatag tgaagaatga tttgatagca
     2161 aaagtggatg aactgacctg tgagaaagat gtgctgcaag gggaattgga ggctgtgaag
     2221 caagccaaac tgaaactaga ggaaaagaac agagaattgg aggaagagct taggaaagct
     2281 cgggcagaag ctgaagatgc aaggcaaaaa gcaaaagatg acgatgatag tgatattccc
     2341 acagcccaga ggaaacggtt tactagagta gaaatggccc gtgttctcat ggagcgaaac
     2401 cagtataaag agagattgat ggagcttcag gaagctgttc gatggacaga gatgattcgg
     2461 gcatcacgag aaaatccagc catgcaggaa aaaaaaaggt caagcatttg gcagtttttc
     2521 agccgacttt tcagctcctc aagtaacacg actaagaagc ctgaaccacc tgttaatctg
     2581 aagtacaatg cacccacgtc tcatgttact ccgtccgtca agaaaagaag cagcacctta
     2641 tctcagctcc ctggggataa gtccaaagcc tttgatttcc ttagtgaaga aactgaagct
     2701 agtttagcct cacgcagaga acaaaagaga gagcagtatc gtcaggtaaa agcacatgtt
     2761 cagaaggaag acggtagagt gcaggctttt ggctggagtc tgcctcagaa gtacaaacag
     2821 gtaaccaatg gtcaaggtga aaataagatg aaaaatttac ctgtgcctgt ctatctcaga
     2881 cctctggatg aaaaagatac atcaatgaag ctgtggtgtg ctgttggagt caatttatct
     2941 ggtgggaaga ccagagatgg tggttctgtt gttggagcaa gtgtatttta caaggatgtt
     3001 gctggtttgg atacagaagg cagtaaacag cgaagtgcct ctcagagtag tttagataag
     3061 ttagatcagg aacttaagga acagcagaag gagttaaaaa atcaagaaga attatccagt
     3121 ctagtttgga tctgtaccag cactcattcg gctacaaaag ttcttattat tgatgctgtt
     3181 caacctggca acatcctaga cagtttcact gtttgcaact ctcatgttct gtgcattgca
     3241 agtgtgccag gtgcacgaga aacagactac cctgcaggag aagatctttc agaatctggt
     3301 caggtagaca aagcatcttt atgtggaagt atgacaagca acagctcagc agagacagac
     3361 agcctgttag gaggcatcac agtggttggt tgttctgcag aaggtgtgac gggagctgcc
     3421 acttccccta gtacaaatgg tgcttctcca gtgatggata aaccaccaga aatggaagca
     3481 gaaaatagtg aggttgatga aaatgttcca acagcagaag aagcaactga agctacagaa
     3541 gggaatgcgg ggtcagctga agacacagtg gacatctccc aaactggcgt ctacacagag
     3601 catgtcttta cagatccttt gggagttcag atcccagaag acctctcccc agtgtatcag
     3661 tcgagcaatg actcagatgc atataaagat caaatatcag tactgccaaa tgaacaagac
     3721 ttggtgagag aagaagccca gaaaatgagt agtcttttac caactatgtg gcttggagct
     3781 caaaatggct gtttgtatgt ccattcatct gtagcccagt ggaggaaatg tctccattcc
     3841 attaaactta aagattcgat tctcagtatt gtacacgtga agggaatcgt gttagtagcc
     3901 ctggctgacg gcacccttgc aatctttcac agaggagtgg atgggcagtg ggatttgtca
     3961 aactatcacc tcttagacct tggacggcct catcattcca tccgttgcat gactgtggta
     4021 catgacaaag tctggtgtgg ctataggaac aaaatctatg tggtgcagcc aaaggccatg
     4081 aaaatagaga aatcttttga tgcacatccc aggaaggaga gccaagtgcg acagcttgcg
     4141 tgggtggggg atggcgtgtg ggtctccatt cgcttggatt ctacgctccg tctctatcat
     4201 gcacacactt atcaacatct acaggatgtg gacattgagc cttatgtaag caaaatgtta
     4261 ggtactggaa aactgggctt ctcttttgtg agaattacag ctcttatggt gtcttgtaat
     4321 cgtttgtggg tggggacagg aaatggtgtc attatctcca tcccattgac agaaacaaat
     4381 aaaacctcag gtgtaccagg aaatcgtcct ggaagtgtaa tccgtgtata tggtgatgaa
     4441 aacagtgata aagtgactcc agggacattt ataccctatt gttcaatggc acatgcacag
     4501 ctttgcttcc atgggcaccg ggatgctgtg aaattctttg tggcagtccc aggtcaagtc
     4561 atcagcccac aaagtagcag tagtggcacg gatctgacgg gtgacaaagc agggccatct
     4621 gcacaggagc ctggtagtca gacgcccttg aagtctatgc ttgtcatcag tggaggagag
     4681 ggctacatcg acttccgaat gggtgatgaa ggtggagaat cagaacttct tggagaggat
     4741 cttccacttg aaccttctgt caccaaagca gaaaggagtc acttgatagt gtggcaagtg
     4801 atgtatggca atgagtgaaa gggcgaattc gacccagctt tcttgtacaa agtggtgatg
     4861 agagaagggc gaattcgaaa gtggtgatcg gtaccttaat taattaaggt accaggtaag
     4921 tgtacccaat tcgccctata gtgagtcgta ttacaattca ctcgatcgcc cttcccaaca
     4981 gttgcgcagc ctgaatggcg aatggagatc caatttttaa gtgtataatg tgttaaacta
     5041 ctgattctaa ttgtttgtgt attttagatt cacagtccca aggctcattt caggcccctc
     5101 agtcctcaca gtctgttcat gatcataatc agccatacca catttgtaga ggttttactt
     5161 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     5221 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5281 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5341 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     5401 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     5461 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     5521 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     5581 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     5641 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     5701 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     5761 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     5821 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5881 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagaa
     5941 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     6001 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     6061 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     6121 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     6181 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     6241 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     6301 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     6361 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     6421 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     6481 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     6541 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     6601 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     6661 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     6721 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     6781 agccggtctt gtcgatcagg atgatctgga cgaagaacat caggggctcg cgccagccga
     6841 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     6901 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     6961 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     7021 tgaagaactt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     7081 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     7141 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     7201 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     7261 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     7321 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     7381 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     7441 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     7501 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     7561 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     7621 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7681 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7741 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7801 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7861 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     7921 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7981 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     8041 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     8101 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     8161 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     8221 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     8281 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     8341 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     8401 tcccctgatt ctgtggataa ccgtattacc gcc
//
