LOCUS       Exported                8142 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 8142)
  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..8142
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          878..2050
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1853..2047
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    14..2050
                     /label=Mac_Q139D_D150N_E258V-HaloTag-ST
     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"
     CDS             1853..2047
                     /codon_start=1
                     /label=Kir2.1_proximal restriction and clustering signal
                     /translation="QSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTEGVIDMRS
                     MSSIDSFISCATDFPEATRF"
     polyA_signal    2171..2395
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2441..2869
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2883..3212
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3063..3198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3221..3820
                     /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    3980..4101
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4150..4166)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4174..4190
                     /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(4198..4228)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4243..4264
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4552..5137)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5308..6168)
                     /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(6169..6273)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6319..6698
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          6974..7990
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     8084..8103
                     /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 tgcaaagtca
     1861 gcctatcctg aacacaaagg aaatggctcc acagtctaag cctcccgaag agcttgagat
     1921 gtccagtatg ccaagtcccg tggctcccct ccctgccagg actgaaggag tgattgacat
     1981 gaggagtatg tcatctattg atagcttcat ctcttgcgca acagatttcc ccgaggctac
     2041 tcgattctaa tgaaagcttg ctaccgagct cggatccact agtccagtgt ggtggaattc
     2101 tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga
     2161 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2221 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2281 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2341 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2401 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2461 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2521 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2581 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2641 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2701 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2761 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2821 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     2881 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2941 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3001 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3061 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3121 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3181 gaggcttttt tggaggccta ggcttttgca aaaagctccc atgaccgagt acaagcccac
     3241 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     3301 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     3361 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     3421 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     3481 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     3541 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     3601 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     3661 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     3721 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     3781 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ttcgaaatga ccgaccaagc
     3841 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     3901 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     3961 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4021 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4081 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4141 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4201 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4261 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4321 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4381 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4441 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4501 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4561 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4621 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4681 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4741 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4801 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4861 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4921 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4981 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5041 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     5101 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5161 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5221 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5281 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5341 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5401 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5461 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5521 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5581 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5641 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5701 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5761 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5821 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5881 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5941 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6001 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6061 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6121 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6181 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6241 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6301 cgtcgacgga tcggggtcga cattgattat tgactagtta ttaatagtaa tcaattacgg
     6361 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     6421 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     6481 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6541 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     6601 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     6661 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
     6721 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
     6781 aattattttg tgcagcgatg ggggcggggg gggggggcgc gcgccaggcg gggcggggcg
     6841 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
     6901 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
     6961 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
     7021 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
     7081 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct
     7141 gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc
     7201 ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg
     7261 cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg
     7321 gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg
     7381 cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca
     7441 accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct
     7501 ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt
     7561 gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc
     7621 ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc
     7681 gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag
     7741 gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga
     7801 aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc
     7861 tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg
     7921 cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt
     7981 tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga
     8041 attcctcgag gaattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     8101 ggccaatgcc ctggctcaca aataccactg agatctgcat ga
//
