LOCUS       Exported                7580 bp DNA     circular SYN 28-JUN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7580)
  AUTHORS   John Grosso
  TITLE     Direct Submission
  JOURNAL   Exported Mar 31, 2022 from SnapGene 5.3.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7580
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     LTR             414..589
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    652..851
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1052..1468
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     CDS             1566..2165
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     enhancer        2340..2643
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2644..2847
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2798..2818
                     /label=Addgene CMV Forward
     promoter        2892..2910
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2929..4224
                     /codon_start=1
                     /label=FLAG-HA-HA-MM.RAP1
                     /translation="MDYKDDDDKLDGGYPYDVPDYAAGAYPYDVPDYALEAGSAEAMDL
                     GKDPNGPTHSSTLFVREDGSAMSFYVRPSSAKRRLSTLILHGGGTVCRVQEPGAVLLAQ
                     PGEALAEASGDFISTQYILDCVDRNEKLDLEAYRLGLTEQASDPKPGASTEGSTEPEPQ
                     PLTGRIAYTDAEDVAILTYVKENARSPSSVTGNALWKAMEKSSLTQHSWQSLKDRYLKH
                     LRGQEHKYLLGNAPVSPSSQKLKRKAEQDPEAADSGEPQNKRAPDLPEEECVKGEIKEN
                     GEADNKLFEEAAPEFGEAVVDESPDFEIHITMCDGDPPTPEEDSETQPDEEEEEPKVST
                     QEVGTAIKVIRQLMEKFNLDLSTVTQALLKNSGELEATSSFLESGRRPDGYPIWCRQDD
                     LDLQKDDDDTKNALVKKFGAQNVARRIEFRKK"
     CDS             2932..2955
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2968..2994
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3004..3030
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     3040..3072
                     /label=mRAP1_BamHI_CDS_F4
     primer_bind     3040..3065
                     /label=Primer 1
     exon            3046..3703
                     /label=EXON1
     exon            3704..3828
                     /label=EXON2
     exon            3829..4221
                     /label=EXON3
     primer_bind     complement(4195..4231)
                     /label=Primer 2
     primer_bind     complement(4195..4224)
                     /label=mRAP1_XhoI_CDS_R1182
     LTR             4319..4912
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     protein_bind    complement(5165..5198)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(5258..5291)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     rep_origin      complement(5582..6170)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6341..7201)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7202..7306)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc
      121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca
      181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
      241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa
      301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac
      361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa
      421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
      481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
      541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
      601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
      661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
      721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
      781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
      841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
      901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
      961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     1441 tccagccctc actccttctc taggcgccgg aattgatccc ccgggctgca ggtcggccgc
     1501 cacgaccggt gccgccacca tcccctgacc cacgcccctg acccctcaca aggagacgac
     1561 cttccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg
     1621 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc
     1681 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     1741 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     1801 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     1861 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     1921 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     1981 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     2041 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     2101 ccgccgacgt cgagtgcccg aaggaccgcg cgacctggtg catgacccgc aagcccggtg
     2161 cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatggct
     2221 ccgaccgaag ccgacccggg cggccccgcc gaccccgcac ccgcccccga ggcccaccga
     2281 ctctagtaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     2341 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     2401 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     2461 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     2521 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     2581 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     2641 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     2701 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     2761 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     2821 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg
     2881 cttatcgaaa ttaatacgac tcactatagg gagacccaag ctgccaccat ggactacaag
     2941 gacgacgatg acaagctcga tggaggatac ccctacgacg tgcccgacta cgccgctgga
     3001 gcatacccct acgacgtgcc cgactacgcc ctcgaggcag gatccgcgga ggcgatggat
     3061 ctgggtaaag accccaatgg gcccactcac tcctccactc tgttcgtgag agaagacggc
     3121 agcgccatgt cgttttacgt gcggcccagc tcggccaagc gccggctgtc gacgctcatc
     3181 ctgcacggcg gcggcaccgt gtgtcgggtg caggaacccg gagccgtgct tctcgcccag
     3241 cccggggagg cgctggccga ggcttcgggc gacttcatct ccacgcagta catcctagac
     3301 tgcgtggatc gcaacgagaa gctggacctg gaggcctatc ggctgggcct gacggagcag
     3361 gcgtccgatc cgaagcccgg ggcttccacc gagggctcca cggaaccgga gccgcagccc
     3421 ctgaccgggc gcatcgccta caccgacgcg gaagatgtgg ccatcctgac ctacgtgaag
     3481 gagaacgccc gttcgcccag ctcggtcaca ggcaatgcct tgtggaaagc gatggagaag
     3541 agctcgctca cgcagcactc ctggcagtcg ctcaaggacc gctacctcaa gcacctacgg
     3601 ggtcaggagc acaagtacct gctcgggaac gccccggtca gcccgtcctc ccagaagctc
     3661 aaacggaagg cggagcagga ccccgaggcc gcggatagcg gagagccaca gaacaagaga
     3721 gcgccagact tgcctgagga ggagtgtgtg aaaggagaga tcaaggagaa tggagaggca
     3781 gacaacaagc tgtttgagga agccgctccg gagttcggag aagccgtggt ggatgagagc
     3841 cctgactttg aaatacatat aacgatgtgt gatggtgatc cacccacacc cgaggaagac
     3901 tcagaaacac agccagacga ggaggaagaa gaaccaaaag tttctacgca agaagtggga
     3961 actgccatta aggtgatccg gcagctaatg gagaagttca acttggatct atcaacagtt
     4021 acacaggcct tgctgaagaa cagtggtgag ctggaggcca cgtcctcctt cttagagtcg
     4081 gggcggagac ccgacggtta tcccatttgg tgcagacaag atgacttaga tttgcagaag
     4141 gacgatgacg acacgaaaaa tgcactggtc aaaaagtttg gagctcagaa cgttgctcgg
     4201 aggattgaat tccgaaagaa ataaaagctt aggtggaatt ctctgactag ctgatcgagc
     4261 atgcatctag agggccgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     4321 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     4381 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
     4441 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     4501 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     4561 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
     4621 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     4681 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     4741 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
     4801 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     4861 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
     4921 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg
     4981 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     5041 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     5101 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
     5161 actgataact tcgtataatg tatgctatac gaagttatta ggtctgaaga ggagtttacg
     5221 tccagccaag ctagggccgc gatccggaac ccttaatata acttcgtata atgtatgcta
     5281 tacgaagtta tcagtactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     5341 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     5401 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5461 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5521 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5581 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5641 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5701 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5761 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5821 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5881 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5941 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6001 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     6061 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6121 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6181 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6241 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6301 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6361 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6421 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6481 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6541 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6601 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     6661 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6721 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6781 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6841 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6901 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     6961 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7021 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7081 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7141 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7201 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7261 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7321 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     7381 gtatcacgag gccctttcgt cttcaagaat tgctagcaat tgctagcaat tgctagcaat
     7441 tcataccaga tcaccgaaaa ctgtcctcca aatgtgtccc cctcacactc ccaaattcgc
     7501 gggcttctgc ctcttagacc actctaccct attccccaca ctcaccggag ccaaagccgc
     7561 ggcccttccg tttctttgct
//
