LOCUS       Exported                5641 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expression of zebrafish GFP-tagged Rab38a in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    EGFP-Rab38a
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5641)
  TITLE     New_for_revision
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5641)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5641
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        25..1002
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1037..1055
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1037..1054
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1110..1134
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1135..1144
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1141..1857
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1141..1857
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1186..1207)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1447..1466)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1794..1815
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1859..1883)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2635..2655)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        complement(2673..2690)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2695..2829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2719..2738)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2773..2792
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2942..2962)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2942..2960)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2981..2997)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2981..2997)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2994..3016)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3005..3021
                     /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(3029..3059)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3074..3095
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3212..3229)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3383..3971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3463..3482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4142..5002)
                     /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     4765..4784
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5003..5107)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5133..5588)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5270..5291)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5482..5501
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcttcgctat tacgccagtc gaccgccaat tcaatatggc gtatatggac tcatgccaat
       61 tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat
      121 tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat
      181 tcaatatggc ggacctggca ctgtgccaac tggggagggg tctacttggc acggtgccaa
      241 gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa
      301 taatggcggc catattggct atatgccagg atcaatatat aggcaatatc caatatggcc
      361 ctatgccaat atggctattg gccaggttca atactatgta ttggccctat gccatatagt
      421 attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat
      481 accatatatg gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct
      541 ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct
      601 cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat
      661 aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac
      721 gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta
      781 atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc
      841 cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt
      901 caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt
      961 gcctaatggg aggtctatat aagcaatgct cgtttaggga accgccattc tgcctgggga
     1021 cgtcggagca agcttgattt aggtgacact atagaataca agctacttgt tctttttgca
     1081 ggatcccatc gattcgaatt caaggatcaa caagtttgta caaaaaagca ggctgccacc
     1141 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1201 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1261 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1321 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     1381 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1441 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1501 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1561 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     1621 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1681 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1741 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1801 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagcac
     1861 ccagctttct tgtacaaagt ggtgctcgac tggcgagtca caagttgtgc tttttccctc
     1921 cgcggggcgt ttctgtttct cccagagtct ctcccagcgg atcacacgcg tccatgttcg
     1981 cctctgtctc acacagccat gcagaaccat cacctgaagg aacatctcta taaaatcctg
     2041 gtgatcgggg acctgggagt ggggaaaacc agcattatca agcgctacgt gcaccagaac
     2101 ttctcaccaa actaccgtgc caccatcggc gtggatttcg cgctcaaagt tctgaatctc
     2161 gaccaggaga ccgttaggct gcagctctgg gacatcgcag gtcaggagcg tttcgggaac
     2221 atgacgcggg tctattacag agaagcaatg ggagccatca ttgtttttga cgtcacgcgg
     2281 ccgtcgacgt ttgaggcggt gacgaagtgg aaggaggact tggatgcaaa gctgagttta
     2341 tccaacggca aacaagtggc ggcggtgctg ctggccaaca aatgtgatca gggccgagac
     2401 gtgctcacca acaacggcat caagatggag cagttctgcc aggagaacgg cttcgtggga
     2461 tggttcgaaa cttcagccaa ggagaatatt aatatagatg aagccgcaca ctgtctggtg
     2521 aagcacatta tagctaatga gaacgatctg ctgcagtcgg aggttgcaga cactatttca
     2581 ccgcagcagg agtccagtcg tggaggaacc tgctcggcct gtttcaggtc ctgacaactt
     2641 tattatacat agttgcactc gagcctctag aactatagtg agtcgtatta cgtagatcca
     2701 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2761 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2821 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     2881 gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt
     2941 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3001 gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag
     3061 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3121 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3181 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3241 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3301 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3361 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3421 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3481 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3541 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3601 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3661 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3721 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3781 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3841 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3901 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3961 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4021 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4081 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4141 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4201 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4261 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4321 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4381 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4441 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4501 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4561 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4621 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4681 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4741 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4801 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4861 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4921 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4981 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5041 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5101 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     5161 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     5221 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     5281 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     5341 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     5401 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5461 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5521 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     5581 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5641 c
//
