LOCUS       Exported                6061 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expression of WT Rab1a in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    PCMV-intron myc Rab1aWT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6061)
  AUTHORS   Dupre DJ, Robitaille M, Ethier N, Villeneuve LR, Mamarbachi AM, 
            Hebert TE
  TITLE     Seven transmembrane receptor core signaling complexes are assembled 
            prior to plasma membrane trafficking.
  JOURNAL   J Biol Chem. 2006 Nov 10;281(45):34561-73. Epub 2006 Sep 7.
  PUBMED    16959776
REFERENCE   2  (bases 1 to 6061)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6061
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..212
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     163..183
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     209..233
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          373..505
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     488..507
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        550..568
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             583..612
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(1273..1291)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1274..1291)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1311..1328)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1317..1541
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1587..2015
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1674..1693)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1884..1905
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2024..2044)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2029..2358
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2209..2344
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2271..2290
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2425..3219
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2479..2498)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3089..3108
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3393..3514
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3430..3449)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3484..3503
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3563..3579)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3563..3579)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3576..3598)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3587..3603
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3611..3641)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3656..3677
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3794..3811)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3965..4553)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4045..4064)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4724..5584)
                     /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     5347..5366
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5585..5689)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt gacgcaaatg ggcggtaggc
      181 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcactagaa
      241 gctttattgc ggtagtttat cacagttaaa ttgctaacgc agtcagtgct tctgacacaa
      301 cagtctcgaa cttaagctgc agtgactctc ttaaggtagc cttgcagaag ttggtcgtga
      361 ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
      421 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
      481 atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag ctcttaaggc
      541 tagagtactt aatacgactc actataggct agcctcgaca tggaacagaa actcatctct
      601 gaagaggatc tgtctagagg atccaccatg tccagcatga atcccgaata tgattattta
      661 ttcaagttac ttctgattgg cgactcaggg gttggaaagt cttgccttct tcttaggttt
      721 gcagatgata catatacaga aagctacatc agcacaattg gtgtggattt caaaataaga
      781 actatagagt tagacgggaa aacaatcaag cttcaaatat gggacacagc aggccaggaa
      841 agatttcgaa caatcacctc cagttattac agaggagccc atggcatcat agttgtgtat
      901 gatgtgacag atcaggagtc cttcaataat gttaaacagt ggctgcagga aatagatcgt
      961 tatgccagtg aaaatgtcaa caaattgttg gtagggaaca aatgtgatct gaccacaaag
     1021 aaagtagtag actacacaac agcgaaggaa tttgctgatt cccttggaat tccgtttttg
     1081 gaaaccagtg ctaagaatgc aacgaatgta gaacagtctt tcatgacgat ggcagctgag
     1141 attaaaaagc gaatgggtcc cggagcaaca gctggtggtg ctgagaagtc caatgttaaa
     1201 attcagagca ctccagtcaa gcagtcaggt ggaggttgct gctaactcga gcatgcatct
     1261 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
     1321 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1381 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1441 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1501 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     1561 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     1621 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1681 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1741 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1801 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1861 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1921 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1981 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     2041 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2101 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2161 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2221 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2281 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2341 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     2401 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     2461 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2521 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2581 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     2641 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     2701 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     2761 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     2821 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     2881 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     2941 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3001 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3061 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3121 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3181 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     3241 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     3301 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3361 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3421 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3481 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     3541 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3601 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3661 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3721 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3781 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3841 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3901 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3961 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4021 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4081 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4141 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4201 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4261 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4321 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4381 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4441 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4501 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4561 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4621 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4681 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4741 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4801 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4861 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4921 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4981 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5041 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5101 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5161 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5221 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5281 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5341 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5401 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5461 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5521 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5581 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5641 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5701 gaaaagtgcc acctgacgtc gacggatcgg gagatcttca atattggcca ttagccatat
     5761 tattcattgg ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc
     5821 tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca tgttggcatt
     5881 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     5941 tggagttccg cgttacataa cttacggtaa atggcccgcc tggcattatg cccagtacat
     6001 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     6061 g
//
