LOCUS       Exported                5246 bp ds-DNA     circular SYN 19-JUN-2015
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5246)
  AUTHORS   David Bryant
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, May 11, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5246
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     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    591
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    592..609
                     /label=MCS
                     /note="multiple cloning site of fluorescent protein 
                     plasmids"
     CDS             622..1164
                     /codon_start=1
                     /product="""human ADP-ribosylation factor 1"", resistant to
                     ARF1_1 (
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                      
                      
                     
                       TRCN0000039876 , 
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                      
                      
                     
                       CAATGACAGAGAGCGTGTGAA )"
                     /label=hARF1-R
                     /translation="MGNIFANLFKGLFGKKEMRILMVGLDAAGKTTILYKLKLGEIVTT
                     IPTIGFNVETVEYKNISFTVWDVGGQDKIRPLWRHYFQNTQGLIFVVDSNDRERVNEAR
                     EELMRMLAEDELRDAVLLVFANKQDLPNAMNAAEITDKLGLHSLRHRNWYIQATCATSG
                     DGLYEGLDWLSNQLRNQK"
     misc_feature    903..923
                     /label=RNAi-R
                     /note="Silent mutations to rend resistant to ARF1_1 
                     (TRCN0000039876, CAATGACAGAGAGCGTGTGAA)."
     misc_feature    1172..1182
                     /label=MCS
                     /note="multiple cloning site of fluorescent protein 
                     plasmids"
     misc_feature    1183..1193
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1201..1911
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     polyA_signal    2034..2155
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2162..2617)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2644..2748
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2750..3107
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2958..3093
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3142..3936
                     /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    4168..4215
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4544..5132
                     /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 gctagcgcta
      601 ccggactcag atctcgagac catggggaac atcttcgcca acctcttcaa gggccttttt
      661 ggcaaaaaag aaatgcgcat cctcatggtg ggcctggatg ctgcagggaa gaccacgatc
      721 ctctacaagc ttaagctggg tgagatcgtg accaccattc ccaccatagg cttcaacgtg
      781 gaaaccgtgg agtacaagaa catcagcttc actgtgtggg acgtgggtgg ccaggacaag
      841 atccggcccc tgtggcgcca ctacttccag aacacacaag gcctgatctt cgtggtggac
      901 agcaacgaca gggaacgggt caacgaggcc cgtgaggagc tcatgaggat gctggccgag
      961 gacgagctcc gggatgctgt cctcctggtg ttcgccaaca agcaggacct ccccaacgcc
     1021 atgaatgcgg ccgagatcac agacaagctg gggctgcact cactacgcca caggaactgg
     1081 tacattcagg ccacctgcgc caccagcggc gacgggctct atgaaggact ggactggctg
     1141 tccaatcagc tccggaacca gaaggcggta ccgcgggccc gggatccacc ggtcgccacc
     1201 atggtgagca agggcgagga ggataacatg gcctctctcc cagcgacaca tgagttacac
     1261 atctttggct ccatcaacgg tgtggacttt gacatggtgg gtcagggcac cggcaatcca
     1321 aatgatggtt atgaggagtt aaacctgaag tccaccaagg gtgacctcca gttctccccc
     1381 tggattctgg tccctcatat cgggtatggc ttccatcagt acctgcccta ccctgacggg
     1441 atgtcgcctt tccaggccgc catggtagat ggctccggct accaagtcca tcgcacaatg
     1501 cagtttgaag atggtgcctc ccttactgtt aactaccgct acacctacga gggaagccac
     1561 atcaaaggag aggcccaggt gaaggggact ggtttccctg ctgacggtcc tgtgatgacc
     1621 aactcgctga ccgctgcgga ctggtgcagg tcgaagaaga cttaccccaa cgacaaaacc
     1681 atcatcagta cctttaagtg gagttacacc actggaaatg gcaagcgcta ccggagcact
     1741 gcgcggacca cctacacctt tgccaagcca atggcggcta actatctgaa gaaccagccg
     1801 atgtacgtgt tccgtaagac ggagctcaag cactccaaga ccgagctcaa cttcaaggag
     1861 tggcaaaagg cctttaccga tgtgatgggc atggacgagc tgtacaagta aagcggccgc
     1921 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1981 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2041 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2101 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     2161 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2221 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2281 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2341 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2401 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2461 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2521 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2581 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     2641 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2701 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     2761 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2821 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2881 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2941 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3001 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3061 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3121 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3181 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3241 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3301 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     3361 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3421 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3481 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3541 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3601 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3661 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3721 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3781 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3841 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3901 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     3961 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4021 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4081 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4141 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     4201 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4261 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     4321 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     4381 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4441 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4501 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4561 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4621 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4681 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4741 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4801 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4861 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4921 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4981 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5041 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5101 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5161 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5221 tggataaccg tattaccgcc atgcat
//
