LOCUS       Exported                5694 bp ds-DNA     circular SYN 20-FEB-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5694)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Aug 22, 2018 from SnapGene 4.2.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5694
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    1
                     /label=Kanamycin
                     /note="Kanamycin"
     misc_feature    1
                     /label=GFP tag
                     /note="GFP tag"
     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"
     misc_feature    601..645
                     /label=NES
                     /note="Value"
     primer_bind     complement(631..657)
                     /label=TgPH1 F
     misc_feature    658..1374
                     /label=EGFP
                     /note="EGFP"
     primer_bind     658..675
                     /label=FPi-F
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     complement(661..678)
                     /label=FPv-R
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     1306..1326
                     /label=A41
     primer_bind     1311..1332
                     /label=EGFP-C
     primer_bind     1355..1374
                     /label=FPv-F
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     complement(1355..1374)
                     /label=FPi-R
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     1375..1439
                     /label=A
     primer_bind     1375..1400
                     /label=Tgph1-R
     misc_feature    1375..1384
                     /label=MCS
                     /note="MCS"
     primer_bind     1381..1413
                     /label=AUX1 ARNO F
     misc_feature    1385..1414
                     /label=5'
                     /note="5'"
     primer_bind     1414..1431
                     /label=ARNO vector F
     gene            1423..1866
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /label=CYTH2
                     /note="cytohesin 2"
                     /db_xref="GeneID:9266"
                     /db_xref="HGNC:HGNC:9502"
                     /db_xref="MIM:602488"
     CDS             1423..1866
                     /codon_start=1
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /product="cytohesin-2 isoform 2"
                     /label=CYTH2
                     /note="isoform 2 is encoded by transcript variant 2; 
                     pleckstrin homology, Sec7 and coiled/coil domains 2 
                     (cytohesin-2); ARF nucleotide-binding site opener; ARF 
                     exchange factor; PH, SEC7 and coiled-coil domain-containing
                     protein 2; pleckstrin homology, Sec7 and coiled-coil 
                     domains 2; CTC-273B12.8"
                     /db_xref="CCDS:CCDS12722.1"
                     /db_xref="GeneID:9266"
                     /db_xref="HGNC:HGNC:9502"
                     /db_xref="MIM:602488"
                     /protein_id="NP_004219.3"
                     /translation="DLTHTFFNPDREGWLLKLGGRVKTWKRRWFILTDNCLYYFEYTTD
                     KEPRGEIPLENLSIREVDDPRKPNCFELYIPNNKGQLIKACKTEADGRVVEGNHMVYRI
                     SAPTQEEKDEWIKSIQAAVSVDPFYEMLAARKKRISVKKKQEQP"
     exon            1423..1477
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /inference="alignment:same species:1.39.8"
     exon            1478..1554
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /inference="alignment:same species:1.39.8"
     exon            1555..1626
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /inference="alignment:same species:1.39.8"
     primer_bind     1563..1590
                     /label=I303E-F
     exon            1627..1781
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /inference="alignment:same species:1.39.8"
     exon            1782..1866
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /inference="alignment:same species:1.39.8"
     misc_feature    1825..1851
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /note="propagated from UniProtKB/Swiss-Prot (Q99418.2); 
                     Region: C-terminal autoinhibitory region. {ECO:0000250}"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     primer_bind     complement(1833..1871)
                     /label=B
     STS             1835..1866
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /standard_name="A005S30"
                     /label=A005S30
                     /db_xref="UniSTS:58772"
     STS             1835..1866
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /standard_name="G32287"
                     /label=G32287
                     /db_xref="UniSTS:116884"
     primer_bind     complement(1849..1881)
                     /label=ARNO R
     primer_bind     complement(1855..1871)
                     /label=AUX1 ARNO R
     primer_bind     1867..1899
                     /label=ARNO vector F
     primer_bind     1882..1907
                     /label=A
     misc_feature    1882
                     /label=5'
                     /note="5'"
     gene            1891..2337
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /label=CYTH2
                     /note="cytohesin 2"
                     /db_xref="GeneID:9266"
                     /db_xref="HGNC:HGNC:9502"
                     /db_xref="MIM:602488"
     CDS             1891..2337
                     /codon_start=1
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /product="cytohesin-2 isoform 2"
                     /label=CYTH2
                     /note="isoform 2 is encoded by transcript variant 2; 
                     pleckstrin homology, Sec7 and coiled/coil domains 2 
                     (cytohesin-2); ARF nucleotide-binding site opener; ARF 
                     exchange factor; PH, SEC7 and coiled-coil domain-containing
                     protein 2; pleckstrin homology, Sec7 and coiled-coil 
                     domains 2; CTC-273B12.8"
                     /db_xref="CCDS:CCDS12722.1"
                     /db_xref="GeneID:9266"
                     /db_xref="HGNC:HGNC:9502"
                     /db_xref="MIM:602488"
                     /protein_id="NP_004219.3"
                     /translation="DLTHTFFNPDREGWLLKLGGRVKTWKRRWFILTDNCLYYFEYTTD
                     KEPRGEIPLENLSIREVDDPRKPNCFELYIPNNKGQLIKACKTEADGRVVEGNHMVYRI
                     SAPTQEEKDEWIKSIQAAVSVDPFYEMLAARKKRISVKKKQEQP"
     exon            1891..1945
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /inference="alignment:same species:1.39.8"
     exon            1946..2022
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /inference="alignment:same species:1.39.8"
     exon            2023..2094
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /inference="alignment:same species:1.39.8"
     primer_bind     2031..2058
                     /label=I303E-F
     exon            2095..2249
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /inference="alignment:same species:1.39.8"
     exon            2250..2337
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /inference="alignment:same species:1.39.8"
     misc_feature    2293..2319
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /note="propagated from UniProtKB/Swiss-Prot (Q99418.2); 
                     Region: C-terminal autoinhibitory region. {ECO:0000250}"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     primer_bind     complement(2301..2390)
                     /label=B
     STS             2303..2337
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /standard_name="A005S30"
                     /label=A005S30
                     /db_xref="UniSTS:58772"
     STS             2303..2337
                     /gene="CYTH2"
                     /gene_synonym="ARNO; CTS18; CTS18.1; cytohesin-2; PSCD2; 
                     PSCD2L; SEC7L; Sec7p-L; Sec7p-like"
                     /standard_name="G32287"
                     /label=G32287
                     /db_xref="UniSTS:116884"
     primer_bind     complement(2317..2334)
                     /label=ARNO R
     primer_bind     complement(2323..2350)
                     /label=AUX1 ARNO R
     misc_feature    2358..2380
                     /label=3'
                     /note="3'"
     polyA_signal    2482..2603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2610..3065)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3127..3155
                     /label=Bacterial
                     /note="Bacterial"
     promoter        3239..3468
                     /label=SV40early
                     /note="SV40early"
     misc_feature    3406..3541
                     /label=SV40
                     /note="SV40"
     rep_origin      3406..3541
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3590..4384
                     /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"
     polyA_signal    4616..4663
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     misc_feature    4620..4638
                     /label=HSV-TK
                     /note="HSV-TK"
     rep_origin      4992..5580
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 gctagccacc
      601 atggctctgc agaaaaagtt ggaagagctt gagctggatg aggcaccggt cgccaccatg
      661 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      721 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      781 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      841 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
      901 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      961 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1021 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1081 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     1141 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1201 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1261 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1321 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga
     1381 ctcagatctc gagctcaagc ttcgaattct gcagtcgaca tggacctgac ccacaccttc
     1441 ttcaacccgg accgggaggg ctggctcctg aagctggggg gccgggtgaa gacgtggaag
     1501 cggcgctggt ttatcctcac agacaactgc ctctactact ttgagtacac cacggacaag
     1561 gagccccgag gagagatccc cctcgagaat ctgagcatcc gagaggtgga cgacccccgg
     1621 aaaccgaact gctttgaact ttacatcccc aacaacaagg ggcagctcat caaagcctgc
     1681 aaaactgagg cggacggccg agtggtggag ggaaaccaca tggtgtaccg gatctcggcc
     1741 cccacgcagg aggagaagga cgagtggatc aagtccatcc aggcggctgt gagtgtggac
     1801 cccttctatg agatgctggc agcgagaaag aagcggattt cagtcaagaa gaagcaggag
     1861 cagcccgggg gatcgggtgg tgtcgacatg gacctgaccc acaccttctt caacccggac
     1921 cgggagggct ggctcctgaa gctggggggc cgggtgaaga cgtggaagcg gcgctggttt
     1981 atcctcacag acaactgcct ctactacttt gagtacacca cggacaagga gccccgagga
     2041 gagatccccc tcgagaatct gagcatccga gaggtggacg acccccggaa accgaactgc
     2101 tttgaacttt acatccccaa caacaagggg cagctcatca aagcctgcaa aactgaggcg
     2161 gacggccgag tggtggaggg aaaccacatg gtgtaccgga tctcggcccc cacgcaggag
     2221 gagaaggacg agtggatcaa gtccatccag gcggctgtga gtgtggaccc cttctatgag
     2281 atgctggcag cgagaaagaa gcggatttca gtcaagaaga agcaggagca gccctgaggt
     2341 accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca taccacattt
     2401 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     2461 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     2521 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2581 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     2641 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     2701 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     2761 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     2821 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     2881 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2941 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     3001 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     3061 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3121 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3181 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3241 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3301 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3361 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3421 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3481 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3541 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3601 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3661 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3721 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     3781 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3841 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3901 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3961 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4021 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4081 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     4141 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4201 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4261 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4321 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4381 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4441 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4501 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     4561 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     4621 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     4681 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     4741 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     4801 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     4861 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4921 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4981 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5041 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5101 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     5161 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5221 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5281 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5341 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5401 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5461 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5521 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5581 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5641 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
