LOCUS       Exported                7809 bp DNA     circular SYN 02-MAY-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7809)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 2, 2021 from SnapGene Viewer 5.3.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7809
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      288..876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     enhancer        1051..1354
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        1355..1558
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     regulatory      1597..1606
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             1603..2319
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 1603 .. 1605 / #05fd14
                        2: 1606 .. 1608 / #05fd14 / 1a
                        3: 1609 .. 2319 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      2368..2377
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             2374..5454
                     /codon_start=1
                     /label=mouse KIF5A
                     /note="color: #993366"
                     /translation="MAETNNECSIKVLCRFRPLNQAEILRGDKFIPIFQGDDSVIIGGK
                     PYVFDRVFPPNTTQEQVYHACAMQIVKDVLAGYNGTIFAYGQTSSGKTHTMEGKLHDPQ
                     LMGIIPRIARDIFNHIYSMDENLEFHIKVSYFEIYLDKIRDLLDVTKTNLSVHEDKNRV
                     PFVKGCTERFVSSPEEILDVIDEGKSNRHVAVTNMNEHSSRSHSIFLINIKQENVETEQ
                     KLSGKLYLVDLAGSEKVSKTGAEGAVLDEAKNINKSLSALGNVISALAEGTKSYVPYRD
                     SKMTRILQDSLGGNCRTTMFICCSPSSYNDAETKSTLMFGQRAKTIKNTASVNLELTAE
                     QWKKKYEKEKEKTKAQKETIAKLEAELSRWRNGENVPETERLAGEDSALGAELCEETPV
                     NDNSSIVVRIAPEERQKYEEEIRRLYKQLDDKDDEINQQSQLIEKLKQQMLDQEELLVS
                     TRGDNEKVQRELSHLQSENDAAKDEVKEVLQALEELAVNYDQKSQEVEEKSQQNQLLVD
                     ELSQKVATMLSLESELQRLQEVSGHQRKRIAEVLNGLMRDLSEFSVIVGNGEIKLPVEI
                     SGAIEEEFTVARLYISKIKSEVKSVVKRCRQLENLQVECHRKMEVTGRELSSCQLLISQ
                     HEAKIRSLTEYMQTVELKKRHLEESYDSLSDELARLQAHETVHEVALKDKEPDTQDAEE
                     VKKALELQMENHREAHHRQLARLRDEINEKQKTIDELKDLNQKLQLELEKLQADYERLK
                     NEENEKSAKLQELTFLYERHEQSKQDLKGLEETVARELQTLHNLRKLFVQDVTTRVKKS
                     AEMEPEDSGGIHSQKQKISFLENNLEQLTKVHKQLVRDNADLRCELPKLEKRLRATAER
                     VKALEGALKEAKEGAMKDKRRYQQEVDRIKEAVRYKSSGKRGHSAQIAKPVRPGHYPAS
                     SPTNPYGTRSPECISYTNNLFQNYQNLHLQAAPSSTSDMYFASSGATSVAPLASYQKAN
                     MDNGNATDINDNRSDLPCGYEAEDQAKLFPLHQETAAS"
     polyA_signal    5587..5708
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(5715..6170)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        6197..6301
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        6303..6660
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      6511..6646
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             6695..7489
                     /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    7721..7768
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /note="color: #a6acb3"
ORIGIN
        1 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
       61 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
      121 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
      181 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
      241 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
      301 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
      361 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
      421 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
      481 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
      541 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
      601 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
      661 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
      721 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
      781 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
      841 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
      901 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
      961 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     1021 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     1081 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     1141 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     1201 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     1261 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     1321 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     1381 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     1441 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     1501 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     1561 tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca ccatggtgag caagggcgag
     1621 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1681 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1741 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     1801 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     1861 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1921 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1981 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     2041 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     2101 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     2161 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     2221 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     2281 gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag atctcgagct
     2341 caagcttcga attctgcagt cgacgccgcc gccatggcgg agactaacaa cgaatgcagc
     2401 atcaaggtgc tttgccgatt tcggcccctg aaccaggccg agattctgcg gggggacaag
     2461 ttcatcccca ttttccaagg ggacgacagc gtcattattg ggggaaagcc atatgtcttt
     2521 gaccgcgtct tccccccaaa caccactcag gagcaggttt accacgcctg tgccatgcag
     2581 atcgtcaaag acgtccttgc tggttacaat ggcacaatct tcgcttatgg acagacatcc
     2641 tcagggaaaa cgcataccat ggaggggaag ctgcacgacc ctcagctgat gggcatcatt
     2701 ccccggatcg ctcgagacat cttcaaccac atctactcca tggatgagaa ccttgaattc
     2761 cacattaagg tatcttactt cgagatttac ctggataaga tccgtgacct tttggatgtg
     2821 accaagacga acctgtccgt gcatgaggac aaaaaccggg tgccgtttgt caagggttgt
     2881 accgaacgct ttgtgtccag cccagaggag attctggatg tgatcgatga ggggaagtcc
     2941 aaccgtcacg tagctgtcac caacatgaac gagcacagtt ctcggagcca cagcatcttc
     3001 ctcatcaaca tcaagcagga gaacgtagag accgagcaga agctcagcgg gaagctgtac
     3061 ctcgtggatc tggccggaag cgagaaggtc agcaagacag gggcagaggg agccgttctg
     3121 gacgaggcaa agaatatcaa caagtcgctg tcggccctgg ggaacgtgat ctctgcactg
     3181 gcagagggca ccaaaagcta cgtgccgtac cgcgacagca aaatgacgag gattctccag
     3241 gactctctgg gagggaactg caggactacc atgttcatct gctgctcgcc gtccagctac
     3301 aatgacgcag agaccaagtc cacgctcatg tttggacagc gggcgaagac catcaagaac
     3361 actgcctcag tgaatctgga gctgactgct gagcagtgga agaagaagta tgagaaggag
     3421 aaggagaaga ccaaggccca gaaggagaca attgcgaagc tagaggctga gcttagccgg
     3481 tggcgcaatg gagagaatgt gcctgagact gagcgcctgg ctggagagga ctcagctctg
     3541 ggagctgagc tctgcgagga gacccctgtg aatgacaact catccattgt ggtacgcatc
     3601 gcacctgagg aaaggcagaa atatgaggaa gagatccgcc gtctctacaa gcagcttgat
     3661 gacaaggatg atgagatcaa ccagcagagc cagctcattg agaagctgaa gcagcagatg
     3721 ctggaccagg aagagctgct cgtgtccact cggggagaca acgagaaggt ccagcgggag
     3781 cttagccacc tgcagtccga gaacgatgct gcgaaggacg aggtgaagga agtgctgcag
     3841 gccctagagg agctggcggt caactacgac cagaagtccc aggaggtgga ggagaagagc
     3901 cagcagaacc agctgctggt ggacgagctg tcccagaaag tggccaccat gctgtccctg
     3961 gagtccgagc tacagcggct ccaggaggtc agtggacacc agcgaaagcg gatcgctgag
     4021 gtgctgaatg ggctgatgag ggacctgagt gagttcagtg tcatcgtggg caacggcgag
     4081 attaagctgc ccgtggagat cagtggggcc atcgaggagg agttcacggt ggcccggctc
     4141 tacatcagca agatcaagtc ggaggtgaag tccgtggtta agcgatgtcg gcagctggag
     4201 aacctccagg tggagtgtca tcgcaagatg gaggtgaccg gtagggagct gtcatcttgc
     4261 caactgctca tctcacagca tgaggccaag atccgttcac tcacggagta catgcagact
     4321 gtggagttga agaaacggca cctggaagag tcctacgact ccctgagcga tgagcttgcc
     4381 aggctccagg cgcacgaaac tgtacacgag gtagctctga aagacaagga gccagacaca
     4441 caggacgcgg aggaggtgaa gaaggccctg gaactacaga tggagaatca ccgtgaggcc
     4501 catcaccggc agctggcccg cctccgggat gagattaatg agaaacagaa aaccattgat
     4561 gagctgaaag acctgaacca gaagctccag ttagagctgg agaagcttca ggccgactat
     4621 gagaggctga agaatgaaga gaacgagaag agcgccaagc tccaggagct gacatttctg
     4681 tatgagcgac atgagcagtc caagcaggac ctcaaggggc tggaggagac agttgcccgt
     4741 gaactccaga ccctccacaa ccttcgcaag ctgttcgttc aagacgtcac gactcgagtc
     4801 aagaaaagtg cagaaatgga gcccgaggac agtgggggga ttcattccca aaagcagaag
     4861 atctcctttc ttgagaacaa cctggaacag cttacaaagg ttcacaaaca gctggtacgt
     4921 gacaatgcag atctgcgttg tgagcttcct aaattggaaa aacgacttcg ggctacggct
     4981 gagagagtta aggccctgga gggtgcactg aaggaggcca aggagggcgc tatgaaggac
     5041 aagcgtagat accagcagga ggtggaccgc atcaaagaag ccgtgcggta caagagctcc
     5101 ggcaagcggg gccattctgc ccagatcgct aagcctgtga ggcctggcca ctatcctgcc
     5161 tcctcaccca ccaaccccta cggcacccgg agccccgagt gtatcagcta caccaacaac
     5221 ctcttccaga actaccagaa cctgcacctg caggctgcgc ctagctccac ttcagatatg
     5281 tactttgcca gcagcggagc cacatctgtt gcccccttgg cttcctacca gaaggccaac
     5341 atggacaatg gaaatgccac agatatcaac gacaacagga gtgacctgcc gtgtggctat
     5401 gaggctgagg accaggccaa gcttttccct ctccaccaag agacagcagc cagctaagga
     5461 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     5521 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     5581 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5641 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5701 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     5761 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     5821 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     5881 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     5941 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     6001 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     6061 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     6121 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     6181 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     6241 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     6301 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     6361 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     6421 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     6481 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     6541 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     6601 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     6661 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     6721 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     6781 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     6841 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     6901 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     6961 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     7021 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     7081 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     7141 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     7201 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     7261 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     7321 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     7381 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     7441 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     7501 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     7561 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     7621 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     7681 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     7741 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     7801 tctgtcgat
//
