LOCUS       Exported                4718 bp ds-DNA     circular SYN 19-JAN-2022
DEFINITION  visualization of the ER.
ACCESSION   .
VERSION     .
KEYWORDS    BFP-KDEL
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4718)
  AUTHORS   Friedman JR, Lackner LL, West M, DiBenedetto JR, Nunnari J, Voeltz 
            GK
  TITLE     ER tubules mark sites of mitochondrial division.
  JOURNAL   Science. 2011 Oct 21;334(6054):358-62. doi: 10.1126/science.1207385.
            Epub 2011 Sep 1.
  PUBMED    21885730
REFERENCE   2  (bases 1 to 4718)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 19, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4718
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      304..892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     793..812
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1067..1370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1371..1574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1525..1545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1657..2352
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     polyA_signal    2512..2633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2549..2568)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2603..2622
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2640..3095)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2777..2798)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2989..3008
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3122..3226
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3228..3585
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3251..3271)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3436..3571
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3498..3517
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3620..4414
                     /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))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3674..3693)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4284..4303
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4602..4621)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4646..4693
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
       61 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
      121 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
      181 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
      241 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
      301 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
      361 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
      421 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
      481 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
      541 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
      601 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
      661 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
      721 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      781 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      841 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      901 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      961 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     1021 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     1081 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     1141 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     1201 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     1261 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     1321 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     1381 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     1441 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     1501 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     1561 atataagcag agctggttta gtgaaccgtc agatccgcta gcatgaagct ctccctggtg
     1621 gccgcgatgc tgctgctgct cagcgcggcg cgggccagcg agctgattaa ggagaacatg
     1681 cacatgaagc tgtacatgga gggcaccgtg gacaaccatc acttcaagtg cacatccgag
     1741 ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc
     1801 cctctcccct tcgccttcga catcctggct actagcttcc tctacggcag caagaccttc
     1861 atcaaccaca cccagggcat ccccgacttc ttcaagcagt ccttccctga gggcttcaca
     1921 tgggagagag tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc
     1981 ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt cacatccaac
     2041 ggccctgtga tgcagaagaa aacactcggc tgggaggcct tcaccgagac gctgtacccc
     2101 gctgacggcg gcctggaagg cagaaacgac atggccctga agctcgtggg cgggagccat
     2161 ctgatcgcaa acatcaagac cacatataga tccaagaaac ccgctaagaa cctcaagatg
     2221 cctggcgtct actatgtgga ctacagactg gaaagaatca aggaggccaa caacgagacc
     2281 tacgtcgagc agcacgaggt ggcagtggcc agatactgcg acctccctag caaactgggg
     2341 cacaagctta ataaagacga gctatgaggt accgcgggcc cgggatccac cggatctaga
     2401 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2461 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2521 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2581 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     2641 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2701 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2761 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2821 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2881 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2941 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3001 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3061 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3121 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3181 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3241 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3301 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3361 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3421 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3481 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3541 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3601 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3661 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3721 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3781 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3841 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3901 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3961 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4021 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4081 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4141 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4201 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4261 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4321 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4381 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4441 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4501 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4561 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4621 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4681 gggtcgtttg ttcataaacg cggggttcgg tcccaggg
//
