LOCUS       Exported                7947 bp ds-DNA     circular SYN 24-JUN-2019
DEFINITION  pcDNA3.1/Puro-CAG-ASAP1 sequence 7389 bps.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7947)
  AUTHORS   Eric Schreiter
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 26, 2019 from SnapGene Viewer 4.3.10
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7947
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          878..1855
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    14..1855
                     /label=Mac_Q139D_D150N_E258V-HaloTag
     CDS             14..877
                     /codon_start=1
                     /product="LeptosphaeriaRh_DpdN_EprV"
                     /label=Mac_Q139D_D150N_E258V
                     /translation="MIVDQFEEVLMKTSQLFPLPTATQSAQPTHVAPVPTVLPDTPIYE
                     TVGDSGSKTLWVVFVLMLIASAAFTALSWKIPVNRRLYHVITTIITLTAALSYFAMATG
                     HGVALNKIVIRTQHDHVPDTYETVYRQVYYARYIDWAITTPLLLLNLGLLAGMSGAHIF
                     MAIVADLIMVLTGLFAAFGSEGTPQKWGWYTIACIAYIFVVWHLVLNGGANARVKGEKL
                     RSFFVAIGAYTLILWTAYPIVWGLADGARKIGVDGVIIAYAVLDVLAKGVFGAWLLVTH
                     ANLRESD"
     CDS             878..1771
                     /codon_start=1
                     /label=HaloI4
                     /label=HaloI4-G&ERexport
                     /translation="IGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYVW
                     RNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHDW
                     GSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQN
                     VFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEEY
                     MDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIGS
                     EIARWLSTLEISGEPTT"
     CDS             1772..1831
                     /codon_start=1
                     /label=TS from Kir2.1
                     /translation="KSRITSEGEYIPLDQIDINV"
     CDS             1832..1852
                     /codon_start=1
                     /label=ER export sequence from Kir2.1
                     /translation="FCYENEV"
     polyA_signal    1976..2200
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2246..2674
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2688..3017
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2868..3003
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3026..3625
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    3785..3906
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3955..3971)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3979..3995
                     /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(4003..4033)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4048..4069
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4357..4942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5113..5973)
                     /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"
     promoter        complement(5974..6078)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6124..6503
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          6779..7795
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7889..7908
                     /label=f.pTriExUP.09.21
ORIGIN
        1 gctagccgcc accatgatcg ttgatcagtt cgaggaggtt ctcatgaaaa catcacagct
       61 cttccctctg ccaactgcaa cacaatctgc ccaacctacc catgttgcac ctgtaccgac
      121 cgtccttcct gacacaccga tttacgagac cgtcggagat agtggttcca aaactctttg
      181 ggtagtattc gtgttgatgc tcatagcatc cgcagctttc acggccttga gctggaagat
      241 acctgtaaac cggaggttgt accacgtaat caccactatc atcacactca cagccgcact
      301 gtcttatttt gccatggcaa ccggccatgg agtcgccctc aacaagatag tcatcaggac
      361 gcaacacgac catgtgccgg acacctatga aaccgtatat aggcaggtat actacgcacg
      421 gtacattgat tgggcaataa caaccccatt gcttctgctg aatctcgggt tgcttgcagg
      481 gatgtcagga gcacacattt ttatggcaat tgttgccgat cttattatgg tcttgacggg
      541 attgttcgct gcctttggca gtgaagggac tcctcaaaaa tggggctggt atacgattgc
      601 ttgcatcgcc tacatctttg ttgtctggca tctcgtactt aacggcggtg ctaacgccag
      661 ggttaaaggg gagaagcttc ggtctttttt cgttgctatc ggagcctaca ccctcatctt
      721 gtggacagct tatcctatcg tttggggcct ggccgacggg gctcgcaaaa ttggcgtaga
      781 cggggtgatt attgcctatg cagttctgga tgtactggct aaaggggtct tcggtgcatg
      841 gctcctggtc actcatgcca atcttcgcga atcagacatc ggtaccggct ttccattcga
      901 cccccattat gtggaagtcc tgggcgagcg catgcactac gtcgatgttg gtccgcgcga
      961 tggcacccct gtgctgttcc tgcacggtaa cccgacctcc tcctacgtgt ggcgcaacat
     1021 catcccgcat gttgcaccga cccatcgctg cattgctcca gacctgatcg gtatgggcaa
     1081 atccgacaaa ccagacctgg gttatttctt cgacgaccac gtccgcttca tggatgcctt
     1141 catcgaagcc ctgggtctgg aagaggtcgt cctggtcatt cacgactggg gctccgctct
     1201 gggtttccac tgggccaagc gcaatccaga gcgcgtcaaa ggtattgcat ttatggagtt
     1261 catccgccct atcccgacct gggacgaatg gccagaattt gcccgcgaga ccttccaggc
     1321 cttccgcacc accgacgtcg gccgcaagct gatcatcgat cagaacgttt ttatcgaggg
     1381 tacgctgccg atgggtgtcg tccgcccgct gactgaagtc gagatggacc attaccgcga
     1441 gccgttcctg aatcctgttg accgcgagcc actgtggcgc ttcccaaacg agctgccaat
     1501 cgccggtgag ccagcgaaca tcgtcgcgct ggtcgaagaa tacatggact ggctgcacca
     1561 gtcccctgtc ccgaagctgc tgttctgggg caccccaggc gttctgatcc caccggccga
     1621 agccgctcgc ctggccaaaa gcctgcctaa ctgcaaggct gtggacatcg gcccgggtct
     1681 gaatctgctg caagaagaca acccggacct gatcggcagc gagatcgcgc gctggctgtc
     1741 gacgctcgag atttccggcg agccaaccac taagagcagg atcaccagcg agggcgagta
     1801 catccccctg gaccagatcg acatcaacgt gttctgctac gagaacgagg tgtaatgaaa
     1861 gcttgctacc gagctcggat ccactagtcc agtgtggtgg aattctgcag atatccagca
     1921 cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     1981 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2041 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2101 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2161 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2221 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2281 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2341 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2401 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2461 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2521 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2581 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2641 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     2701 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2761 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2821 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2881 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2941 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3001 gcctaggctt ttgcaaaaag ctcccatgac cgagtacaag cccacggtgc gcctcgccac
     3061 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     3121 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     3181 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     3241 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     3301 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     3361 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     3421 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     3481 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     3541 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     3601 catgacccgc aagcccggtg cctgattcga aatgaccgac caagcgacgc ccaacctgcc
     3661 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3721 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3781 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3841 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3901 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     3961 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4021 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4081 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4141 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4201 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4261 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4321 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4381 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4441 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4501 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4561 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4621 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4681 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4741 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4801 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4861 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     4921 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4981 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5041 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5101 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5161 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5221 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5281 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5341 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5401 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5461 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5521 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5581 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5641 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5701 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5761 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5821 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5881 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5941 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6001 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6061 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     6121 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     6181 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     6241 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     6301 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     6361 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     6421 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     6481 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
     6541 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
     6601 cgatgggggc gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
     6661 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
     6721 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
     6781 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
     6841 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
     6901 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
     6961 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt
     7021 gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg
     7081 cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg
     7141 cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg
     7201 cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc
     7261 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg
     7321 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
     7381 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg
     7441 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca
     7501 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc
     7561 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     7621 ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg
     7681 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     7741 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
     7801 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcc tcgaggaatt
     7861 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     7921 tcacaaatac cactgagatc tgcatga
//
