LOCUS       Exported                6188 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pT7-EGFP-C1-HsRCK
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6188)
  AUTHORS   Tritschler F, Braun JE, Motz C, Igreja C, Haas G, Truffault V, 
            Izaurralde E, Weichenrieder O
  TITLE     DCP1 forms asymmetric trimers to assemble into active mRNA decapping
            complexes in metazoa.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Dec 22. 106(51):21591-6.
  PUBMED    19966221
REFERENCE   2  (bases 1 to 6188)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6188
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     671..690
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        671..689
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      717..726
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             723..1439
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             723..1439
                     /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(768..789)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1029..1048)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1376..1397
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3034..3155
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3071..3090)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3125..3144
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3162..3617)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3299..3320)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3511..3530
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3644..3748
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3750..4107
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3773..3793)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3958..4093
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4020..4039
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4142..4936
                     /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(4196..4215)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4806..4825
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5124..5143)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5168..5215
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5544..6132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6033..6052
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
      661 ttatcgaaat taatacgact cactataggg agacccaagc tctagcgcta ccggtcgcca
      721 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      781 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      841 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      901 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
      961 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1021 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1081 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1141 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     1201 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1261 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1321 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1381 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     1441 ccggactcag atctatgggt ctgtccagtc aaaatggtca gctgagaggc cctgtgaaac
     1501 ccactggtgg ccctggagga gggggcacac agacacagca acagatgaac cagctgaaaa
     1561 acaccaacac aatcaataat ggcactcagc agcaagcaca gagtatgacc accactatta
     1621 aacctggtga tgactggaaa aagactttaa aactccctcc aaaggatcta agaatcaaaa
     1681 cttcggatgt gacctccaca aaaggaaatg agtttgaaga ttactgtttg aaacgggagt
     1741 tactgatggg aatttttgaa atgggctggg aaaagccatc tcctattcag gaggagagca
     1801 ttcccattgc tttatctggt agggatatct tagctagagc aaaaaatgga acaggcaaga
     1861 gcggtgccta cctcattccc ttacttgaac ggctagacct gaagaaggac aatatacaag
     1921 caatggtgat tgttcccact agagaacttg ctctacaggt cagtcaaatt tgcatccagg
     1981 tcagcaaaca catgggaggg gccaaagtga tggcaaccac aggaggaacc aatttacgag
     2041 atgacataat gaggcttgat gatacagtgc acgtggtgat tgctacccct gggagaatcc
     2101 tggatcttat taagaaagga gtagcaaagg ttgatcatgt ccagatgata gtattggatg
     2161 aggcagataa gttgctgtca caggattttg tgcagataat ggaggatatt attctcacgc
     2221 tacctaaaaa caggcagatt ttactatatt ccgctacttt ccctcttagt gtacagaagt
     2281 tcatgaattc ccatttgcag aaaccctatg agattaacct gatggaggaa ctaactctga
     2341 agggagtaac ccagtactac gcatatgtaa ctgagcgcca aaaagtacac tgcctcaaca
     2401 cacttttctc caggcttcag ataaaccagt cgatcatttt ctgtaactcc tctcagcgag
     2461 ttgaattgct agccaagaag atttctcaac tgggttattc ttgcttctat attcatgcta
     2521 aaatgaggca ggaacatcga aatcgtgtat ttcatgattt ccgaaatggc ttatgccgca
     2581 atcttgtttg cactgatctg tttacccgag gtattgatat acaagctgtg aatgtggtaa
     2641 taaactttga tttcccaaag ctggcagaga cctatctcca tcgtattgga agatcaggtc
     2701 gctttggtca tcttggctta gccatcaact tgatcacata tgatgatcgc ttcaacctga
     2761 aaagtattga ggagcagctg ggaacagaaa ttaaacctat tccgagcaac attgataaga
     2821 gcctgtatgt ggcagaatac cacagcgagc ctgtagaaga tgagaaacct taacaattct
     2881 gcagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc
     2941 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3001 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3061 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3121 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     3181 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3241 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3301 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3361 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3421 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3481 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3541 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3601 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3661 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3721 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3781 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3841 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3901 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3961 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4021 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4081 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     4141 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4201 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4261 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4321 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4381 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4441 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4501 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4561 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4621 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4681 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4741 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4801 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4861 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4921 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4981 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5041 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5101 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5161 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     5221 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     5281 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     5341 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     5401 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5461 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5521 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5581 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5641 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5701 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5761 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5821 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5881 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5941 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6001 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6061 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6121 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6181 tttgctca
//
