LOCUS       Exported                7788 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 7788)
  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..7788
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          719..1696
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    14..1696
                     /label=Ace1_Q89D_D100N_E206V-HaloTag-ST
     CDS             14..718
                     /codon_start=1
                     /product="AcetabulariaRh_DpdN_EprV"
                     /label=AcetabulariaRh_DQ89D_100N_E206V
                     /translation="MSNPNPFQTTLGTDAQWVVFAVMALAAIVFSIAVQFRPLPLRLTY
                     YVNIAICTIAATAYYAMAVNGGDNKPTAGTGADERQVIYARYIDWVFTTPLLLLNLVLL
                     TNMPATMIAWIMGADIAMIAFGIIGAFTVGSYKWFYFVVGCIMLAVLAWGMINPIFKEE
                     LQKHKEYTGAYTTLLIYLIVLWVIYPIVWGLGAGGHIIGVDVVIIAMGVLDLLAKPLYA
                     IGVLITVEVVYGK"
     CDS             719..1612
                     /codon_start=1
                     /label=HaloI4
                     /label=HaloI4-G&ERexport
                     /translation="IGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYVW
                     RNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHDW
                     GSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQN
                     VFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEEY
                     MDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIGS
                     EIARWLSTLEISGEPTT"
     CDS             1613..1672
                     /codon_start=1
                     /label=TS from Kir2.1
                     /translation="KSRITSEGEYIPLDQIDINV"
     CDS             1673..1693
                     /codon_start=1
                     /label=ER export sequence from Kir2.1
                     /translation="FCYENEV"
     polyA_signal    1817..2041
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2087..2515
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2529..2858
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2709..2844
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2867..3466
                     /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    3626..3747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3796..3812)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3820..3836
                     /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(3844..3874)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3889..3910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4198..4783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4954..5814)
                     /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(5815..5919)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5965..6344
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          6620..7636
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7730..7749
                     /label=f.pTriExUP.09.21
ORIGIN
        1 gctagccgcc accatgagca atccaaatcc ctttcagact actcttggca cggacgccca
       61 gtgggttgtc tttgccgtaa tggcactggc agccatagtg tttagtatag cagttcagtt
      121 tcggccactc ccgctgcgcc tcacttacta cgtcaacatt gctatttgta caatagccgc
      181 aacagcttac tatgccatgg ccgttaatgg cggagacaat aaaccgacag ctgggacggg
      241 ggcagacgaa cgccaggtta tttacgctcg ctatatcgac tgggtgttta ccacaccact
      301 cctcttgctt aatctggtgt tgttgactaa catgccagcc actatgatcg cttggattat
      361 gggagcagac atcgctatga tcgcttttgg aatcatcggt gctttcaccg tcggctctta
      421 caagtggttt tacttcgtag tggggtgcat aatgctcgca gtccttgctt gggggatgat
      481 aaaccccata ttcaaagagg aattgcagaa acacaaagag tacacagggg cctacacgac
      541 tctgctcata tatctcatag tcttgtgggt tatatatcca attgtctggg gtctcggagc
      601 aggcgggcac attataggag tggatgtagt gattatcgca atgggtgttc tggacctgct
      661 tgccaagccg ctttacgcta taggggttct cataacggtt gaggttgttt acggaaagat
      721 cggtaccggc tttccattcg acccccatta tgtggaagtc ctgggcgagc gcatgcacta
      781 cgtcgatgtt ggtccgcgcg atggcacccc tgtgctgttc ctgcacggta acccgacctc
      841 ctcctacgtg tggcgcaaca tcatcccgca tgttgcaccg acccatcgct gcattgctcc
      901 agacctgatc ggtatgggca aatccgacaa accagacctg ggttatttct tcgacgacca
      961 cgtccgcttc atggatgcct tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat
     1021 tcacgactgg ggctccgctc tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa
     1081 aggtattgca tttatggagt tcatccgccc tatcccgacc tgggacgaat ggccagaatt
     1141 tgcccgcgag accttccagg ccttccgcac caccgacgtc ggccgcaagc tgatcatcga
     1201 tcagaacgtt tttatcgagg gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt
     1261 cgagatggac cattaccgcg agccgttcct gaatcctgtt gaccgcgagc cactgtggcg
     1321 cttcccaaac gagctgccaa tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga
     1381 atacatggac tggctgcacc agtcccctgt cccgaagctg ctgttctggg gcaccccagg
     1441 cgttctgatc ccaccggccg aagccgctcg cctggccaaa agcctgccta actgcaaggc
     1501 tgtggacatc ggcccgggtc tgaatctgct gcaagaagac aacccggacc tgatcggcag
     1561 cgagatcgcg cgctggctgt cgacgctcga gatttccggc gagccaacca ctaagagcag
     1621 gatcaccagc gagggcgagt acatccccct ggaccagatc gacatcaacg tgttctgcta
     1681 cgagaacgag gtgtaatgaa agcttgctac cgagctcgga tccactagtc cagtgtggtg
     1741 gaattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc
     1801 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1861 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1921 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1981 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2041 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2101 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2161 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2221 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2281 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2341 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2401 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2461 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2521 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2581 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2641 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2701 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2761 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     2821 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccatga ccgagtacaa
     2881 gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc
     2941 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg
     3001 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     3061 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     3121 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     3181 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     3241 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     3301 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     3361 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     3421 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgattcg aaatgaccga
     3481 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3541 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3601 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     3661 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3721 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     3781 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3841 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3901 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3961 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4021 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4081 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4141 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4201 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4261 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4321 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4381 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4441 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4501 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4561 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4621 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4681 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     4741 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4801 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4861 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4921 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4981 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5041 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5101 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5161 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5221 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5281 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5341 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5401 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5461 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5521 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5581 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5641 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5701 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5761 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5821 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5881 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5941 acctgacgtc gacggatcgg ggtcgacatt gattattgac tagttattaa tagtaatcaa
     6001 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     6061 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     6121 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     6181 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     6241 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     6301 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
     6361 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
     6421 ttttttaatt attttgtgca gcgatggggg cggggggggg gggcgcgcgc caggcggggc
     6481 ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag
     6541 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
     6601 gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc
     6661 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
     6721 gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc
     6781 ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag
     6841 cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct
     6901 gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg
     6961 cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa
     7021 aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg
     7081 gctgcaaccc cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc
     7141 ggggctccgt acggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt
     7201 gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg
     7261 cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt gccttttatg
     7321 gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa atctgtgcgg agccgaaatc
     7381 tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag
     7441 gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct
     7501 ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg
     7561 gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt
     7621 cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg
     7681 caaagaattc ctcgaggaat tcactcctca ggtgcaggct gcctatcaga aggtggtggc
     7741 tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctgcatga
//
