LOCUS       Exported                5471 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    prLCA-Citrine
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5471)
  AUTHORS   Offterdinger M, Bastiaens PI
  TITLE     Prolonged EGFR signaling by ERBB2-mediated sequestration at the 
            plasma membrane.
  JOURNAL   Traffic. 2008 Jan;9(1):147-55. Epub 2007 Nov 13.
  PUBMED    17956594
REFERENCE   2  (bases 1 to 5471)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5471
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    917..1038
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(954..973)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1008..1027
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1045..1500)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1182..1203)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1394..1413
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1527..1631
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1633..1990
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(1656..1676)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1841..1976
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1903..1922
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2025..2819
                     /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(2079..2098)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2689..2708
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3007..3026)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3051..3098
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3427..4015
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3916..3935
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4190..4493
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4494..4697
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4648..4668
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4736..4745
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4742..5458
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             4742..5458
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4787..4808)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5048..5067)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5395..5416
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ctcgaggcat ggctgagttg gatccatttg gcgctcccgc cggtgcgccc ggtggccccg
       61 cgctcgggaa cggggtggcg ggcgccggcg aggaagaccc ggccgcggcc ttcttggcgc
      121 agcaggagag tgagattgcg ggcatagaga acgacgaggc cttcgccatc ctagacggcg
      181 gcgcccccgg gccgcaggca cacggcgagc cgccgggggg tcctgatgcg gttgatggag
      241 tgatgaatgg cgaatactac caggagagca atggtccaac agacagttac gcagccattt
      301 cagaagtgga tcggttgcag tcagagcctg aaagtatccg taagtggaga gaagagcaga
      361 cagagcgcct ggaagccctc gatgccaatt ctcggaagca ggaagcagag tggaaagaaa
      421 aggcagtgaa ggagctggaa gagtggtacg caaggcagga tgagcagcta cagaagacaa
      481 aagccagcaa cagggtggca gatgaagctt tctacaaaca acccttcgct gacgtgattg
      541 gttatgtcac aaacataaac catccttgct acagcctaga acaggcagca gaagaagcct
      601 ttgtaaacga cattgacgag tcatccccag gcactgagtg ggagcgggtg gcccgcctgt
      661 gtgactttaa ccccaagtcc agcaaacagg ccaaagatgt ctctcgcatg cgctcagtcc
      721 tcatctccct gaagcaggcg cccctggtgc attgaagaat tctgcagtcg acggtaccgc
      781 gggcccggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
      841 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
      901 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
      961 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1021 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     1081 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     1141 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     1201 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     1261 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     1321 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     1381 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     1441 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     1501 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1561 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1621 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     1681 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     1741 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     1801 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     1861 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     1921 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     1981 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     2041 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     2101 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     2161 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     2221 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     2281 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     2341 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     2401 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     2461 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     2521 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     2581 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     2641 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     2701 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     2761 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     2821 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     2881 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     2941 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     3001 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     3061 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3121 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     3181 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     3241 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     3301 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3361 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3421 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3481 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3541 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3601 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3661 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3721 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3781 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3841 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3901 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3961 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4021 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4081 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     4141 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     4201 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     4261 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4321 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4381 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     4441 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     4501 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     4561 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     4621 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     4681 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggtcgccac
     4741 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     4801 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     4861 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     4921 cctcgtgacc accttcggct acggcctgat gtgcttcgcc cgctaccccg accacatgaa
     4981 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     5041 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     5101 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     5161 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     5221 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     5281 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     5341 ctacctgagc taccagtcca aactgagcaa agaccccaac gagaagcgcg atcacatggt
     5401 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc
     5461 cggactcaga t
//
