LOCUS       Exported                6579 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  To express in mammalian cells the endocytic protein Rab7a fused to 
            mEmerald by transfection or by generating AAV virions..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-UbC-mEmerald Rab7a
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6579)
  AUTHORS   Pantelis Tsoulfas
  TITLE     Eucaryotic expression plasmids for expression and visualization of 
            early endocytic markers Rab5 and Rab7
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6579)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6579
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        152..551
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1121..1140
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     primer_bind     1837..1856
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1895..1904
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1901..2617
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1946..1967)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2207..2226)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2554..2575
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3326..3802
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3525..3544)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3842..3982
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3842..3971
                     /label=AAV2 ITR
     rep_origin      4057..4512
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4144..4163)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4354..4375
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4529..4548)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4648..4670
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4708..4726)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4794..4898
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4899..5759
                     /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"
     primer_bind     complement(5117..5136)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5930..6518
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6419..6438
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaaac tggcctccgc gccgggtttt ggcgcctccc
      181 gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag acgaagggcg cagcgagcgt
      241 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
      301 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
      361 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
      421 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
      481 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
      541 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
      601 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
      661 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
      721 gaatagaacc ttcgctaatg cgggaaagct cttattcggg tgagatgggc tggggcacca
      781 tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc
      841 gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc
      901 cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccacctg
      961 ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag
     1021 gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag
     1081 tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact
     1141 atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa
     1201 tcatttgggt caatatgtaa ttttcagtgt tagactagta aattgtccgc taaattctgg
     1261 ccgtttttgg cttttttgtt agacgaagct aaggcgcgcc tgagaacttc agggtgagtt
     1321 tggggaccct tgattgttct ttctttttcg ctattgtaaa attcatgtta tatggagggg
     1381 gcaaagtttt cagggtgttg tttagaatgg gaagatgtcc cttgtatcac catgcatgga
     1441 ccctcatgat aattttgttt ctttcacttt ctactctgtt gacaaccatt gtctcctctt
     1501 attttctttt cattttctgt aactttttcg ttaaacttta gcttgcattt gtaacgaatc
     1561 tagacaatca gggtatatta tattgtactt cagcacagtt ttagagaaca attgttataa
     1621 ttaaatgata aggtagaata tttctgcata taaattctgg ctggcgtgga aatattctta
     1681 ttggtagaaa caactacatc ctggtcatca tcctgccttt ctctttatgg ttacaatgat
     1741 atacactgtt tgagatgagg ataaaatact ctgagtccaa accgggcccc tctgctaacc
     1801 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
     1861 ctcatcaagc ttggatcccc tagcgctacc ggtcgccacc atggtgagca agggcgagga
     1921 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1981 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     2041 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttgaccta
     2101 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2161 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     2221 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2281 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2341 cagccacaag gtctatatca ccgccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     2401 gacccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2461 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa
     2521 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2581 cgccgggatc actctcggca tggacgagct gtacaagtcc ggactcagat ctcgagccat
     2641 gacctctagg aagaaagtgt tgctgaaggt tatcatcctg ggagattctg gagtcgggaa
     2701 gacatcactc atgaaccagt atgtgaataa gaaattcagc aatcagtaca aagccacaat
     2761 aggagctgac tttctgacca aggaggtgat ggtggatgac aggctagtca caatgcagat
     2821 atgggacaca gcaggacagg aacggttcca gtctctcggt gtggccttct acagaggtgc
     2881 agactgctgc gttctggtat ttgatgtgac tgcccccaac acattcaaaa ccctagatag
     2941 ctggagagat gagtttctca tccaggccag tccccgagat cctgaaaact tcccatttgt
     3001 tgtgttggga aacaagattg acctcgaaaa cagacaagtg gccacaaagc gggcacaggc
     3061 ctggtgctac agcaaaaaca acattcccta ctttgagacc agtgccaagg aggccatcaa
     3121 cgtggagcag gcgttccaga cgattgcacg gaatgcactt aagcaggaaa cggaggtgga
     3181 gctgtacaac gaatttcctg aacctatcaa actggacaag aatgaccggg ccaaggcctc
     3241 ggcagaaagc tgcagttgct gaggatcggg ctgcaggaat tcgatatctc gagtctagag
     3301 ttaacggcca tattggccaa tctacgggtg gcatccctgt gacccctccc cagtgcctct
     3361 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     3421 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     3481 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     3541 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     3601 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     3661 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     3721 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     3781 cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg
     3841 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3901 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     3961 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     4021 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     4081 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     4141 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4201 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4261 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4321 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4381 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     4441 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4501 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4561 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4621 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4681 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4741 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4801 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4861 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4921 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4981 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5041 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5101 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5161 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5221 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5281 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5341 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5401 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5461 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5521 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5581 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5641 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5701 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5761 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5821 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5881 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5941 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6001 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6061 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6121 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6181 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6241 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6301 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6361 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6421 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6481 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6541 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
