LOCUS       Exported                5240 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    CAV1-K*R-mEGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5240)
  AUTHORS   Hayer A, Stoeber M, Ritz D, Engel S, Meyer HH, Helenius A
  TITLE     Caveolin-1 is ubiquitinated and targeted to intralumenal vesicles in
            endolysosomes for degradation.
  JOURNAL   J Cell Biol. 2010 Nov 1. 191(3):615-29.
  PUBMED    21041450
REFERENCE   2  (bases 1 to 5240)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5240
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1180..1189
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1186..1905
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1231..1252)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1492..1511)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2028..2149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2065..2084)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2119..2138
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2156..2611)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2293..2314)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2505..2524
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2638..2742
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2744..3101
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2767..2787)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2952..3087
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3014..3033
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3136..3930
                     /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(3190..3209)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3800..3819
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4118..4137)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4162..4209
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4538..5126
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5027..5046
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttacc atgtctgggg gccgctacgt agactcggag
      661 ggacatctct acaccgttcc catccgggaa cagggcaaca tctaccgccc caacaaccgc
      721 gccatggcag acgagctgag cgagcgccaa gtgtacgacg cgcacacccg cgagatcgac
      781 ctggtcaacc gcgaccctcg ccacctcaac gatgacgtgg tccgcattga ctttgaagat
      841 gtgattgcag aaccagaagg gacacacagt tttgacggca tttggcgcgc cagcttcacc
      901 accttcactg tgacgcgcta ctggttttac cgcttgctgt ctgccctctt tggcatcccg
      961 atggcactca tctggggcat ttacttcgcc attctctctt tcctgcacat ctgggcagtt
     1021 gtaccatgca ttcgcagctt cctgattgag attcagtgca tcagccgtgt ctattccatc
     1081 tacgtccaca ccgtctgtga cccactcttt gaagctgttg ggcgcatatt cagcaatgtc
     1141 cgcatcaact tgcagcgcga aataggggat ccaccggtcg ccaccatggt gagcaagggc
     1201 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1261 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1321 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1381 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     1441 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1501 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1561 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1621 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     1681 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1741 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     1801 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtccttaa ggagttcgtg
     1861 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct
     1921 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     1981 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2041 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2101 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     2161 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2221 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2281 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2341 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2401 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2461 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2521 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2581 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2641 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2701 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     2761 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2821 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2881 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2941 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3001 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3061 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3121 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3181 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3241 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3301 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3361 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3421 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3481 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3541 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3601 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3661 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3721 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3781 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3841 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3901 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3961 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4021 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4081 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4141 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4201 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4261 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4321 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4381 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4441 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4501 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4561 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4621 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4681 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4741 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4801 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4861 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4921 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4981 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5041 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5101 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5161 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5221 accgtattac cgccatgcat
//
