LOCUS       Exported                6527 bp ds-DNA     circular SYN 14-SEP-2020
DEFINITION  Plasmid for expressing pIgH-ESCargo(FTV) in cultured mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6527)
  AUTHORS   Glick Lab
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Sep 17, 2020 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Alias: pEF-FTVNTT-DsRed-Express2-FKBP-LV(C22V)
FEATURES             Location/Qualifiers
     source          1..6527
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     rep_origin      96..231
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             271..1065
                     /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="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    1239..1360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1564..2152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2323..3183)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3184..3288)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3364..4526
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3595..4517
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     regulatory      4534..4543
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     sig_peptide     join(4540..4584,4667..4678)
                     /product="modified signal sequence from the mouse 
                     immunoglobulin heavy chain variable region"
                     /label=IgH signal sequence
     CDS             4679..4696
                     /codon_start=1
                     /product="tripeptide ER export signal followed by 
                     tripeptide N-glycosylation signal"
                     /label=FTVNTT
                     /translation="FTVNTT"
     CDS             4697..5368
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="DSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAKL
                     QVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHKA
                     LKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHHL
                     FQ"
     CDS             5381..5701
                     /codon_start=1
                     /product="cysteine-free improved reversibly dimerizing 
                     mutant of FK506-binding protein FKBP12 (Barrero et al., 
                     2016; Casler et al., 2019)"
                     /label=FKBP-LV(C22V)
                     /translation="GVQVETISPGDGRTFPKRGQTVVVHYTGMLEDGKKLDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGVIPPHATLVFDVELL
                     KLE"
     polyA_signal    5731..5955
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6001..6429
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        join(6443..6527,1..245)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
       61 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      121 ccgcccattc tccgccccta ggctgactaa ttttttttat ttatgcagag gccgaggccg
      181 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      241 gcaaaaagct ccccccccgg gaggtccaca atggttgaac aagatggatt gcacgcaggt
      301 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
      361 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
      421 accgacctgt ccggtgccct gaatgaactc caggacgagg cagcgcggct atcgtggctg
      481 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
      541 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
      601 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
      661 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
      721 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
      781 ttcgccaggc tcaaggcgcg tatgcccgac ggcgaggatc tcgtcgtgac tcatggcgat
      841 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
      901 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
      961 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     1021 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     1081 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     1141 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     1201 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     1261 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1321 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     1381 tctttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1441 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1501 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1561 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1621 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1681 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1741 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1801 ctccaagctg ggctgtatgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1861 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1921 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1981 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2041 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2101 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2161 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2221 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2281 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2341 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2401 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2461 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2521 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2581 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2641 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2701 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2761 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2821 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2881 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     2941 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3001 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3061 tcgggcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3121 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3181 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3241 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3301 aaaagtgcca cctgacgtca gatcgacgga tcgggagatc ggtaccgaat tcaagcttcg
     3361 tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     3421 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
     3481 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     3541 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
     3601 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
     3661 attacttcca cctggctcca gtacgtgatt cttgatcccg agctggagcc aggggcgggc
     3721 cttgcgcttt aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
     3781 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
     3841 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
     3901 taaatgcggg ccaggatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
     3961 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     4021 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     4081 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     4141 aaagatggcc gcttcccggc cctgctccag ggggctcaaa atggaggacg cggcgctcgg
     4201 gagagcgggc gggtgagtca cccacacaaa ggaaaggggc ctttccgtcc tcagccgtcg
     4261 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctggagct
     4321 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     4381 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     4441 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     4501 tttttcttcc atttcaggtg tcgtgaacac gtggccacca tgggatggag ctgtatcatc
     4561 ctcttcttgg tagcaacagc tacaggtaag gggttaacag tagcaggctt gaggtctgga
     4621 catatatatg ggtgacaatg acatccactt tgcctttctc tccacaggcg cgcactcctt
     4681 taccgtgaac accaccgata gcactgagaa cgtcatcaag cccttcatgc gcttcaaggt
     4741 gcacatggag ggctcagtaa atggtcatga gttcgagatc gagggcgagg gcgagggcaa
     4801 gccctacgag ggcacccaga ccgccaagct gcaggtgacc aagggcggcc ccctgccctt
     4861 cgcctgggac atcctgtccc cccagttcca gtacggctcc aaggtgtacg tgaagcaccc
     4921 cgccgacatc cccgactaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     4981 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     5041 caccttcatc taccacgtga agttcatcgg cgtgaacttc ccctccgacg gccccgtaat
     5101 gcagaagaag actctgggct gggagccctc caccgagcgc ctgtaccccc gcgacggcgt
     5161 gctgaagggc gagatccaca aggcgctgaa gctgaagggc ggcggccact acctggtgga
     5221 gttcaagtca atctacatgg ccaagaagcc cgtgaagctg cccggctact actacgtgga
     5281 ctccaagctg gacatcacct cccacaacga ggactacacc gtggtggagc agtacgagcg
     5341 cgccgaggcc cgccaccacc tgttccaggg aggatccaga ggagtgcagg tggaaaccat
     5401 ctccccagga gacgggcgca ccttccccaa gcgcggccag accgttgtgg tgcactacac
     5461 cgggatgctt gaagatggaa agaaactaga ttcctcccgg gacagaaaca agccctttaa
     5521 gtttatgcta ggcaagcagg aggtgatccg aggctgggaa gaaggggttg cccagatgag
     5581 tgtgggtcag agagccaaac tgactatatc tccagattat gcctatggtg ccactgggca
     5641 cccaggcgtt atcccaccac atgccactct cgtcttcgat gtggagcttc taaaactgga
     5701 atagtctaga agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     5761 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     5821 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     5881 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     5941 gcccgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     6001 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     6061 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     6121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     6181 gtgctttacg gcacctctcc cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     6241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     6301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     6361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     6421 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     6481 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaacca
//
