LOCUS       Exported                8010 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 8010)
  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..8010
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          746..1918
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1721..1915
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    14..1918
                     /label=QuaAr3_Q95D_H106N_E214V-HaloTag-ST
     CDS             14..745
                     /codon_start=1
                     /product="QuasAr3"
                     /label=QuaAr3_Q95D_H106N_E214V
                     /translation="MVSIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT
                     DKDAREYYAVTILVSGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYADWLFTTPLLL
                     LNLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA
                     TSLRSAARERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIVTLLFMVLD
                     VTAKVGFGFILLRSRAILGDTE"
     CDS             746..1639
                     /codon_start=1
                     /label=HaloI4
                     /label=HaloI4-G&ERexport
                     /translation="IGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYVW
                     RNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHDW
                     GSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQN
                     VFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEEY
                     MDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIGS
                     EIARWLSTLEISGEPTT"
     CDS             1640..1699
                     /codon_start=1
                     /label=TS from Kir2.1
                     /translation="KSRITSEGEYIPLDQIDINV"
     CDS             1700..1720
                     /codon_start=1
                     /label=ER export sequence from Kir2.1
                     /translation="FCYENEV"
     CDS             1721..1915
                     /codon_start=1
                     /label=Kir2.1_proximal restriction and clustering signal
                     /translation="QSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTEGVIDMRS
                     MSSIDSFISCATDFPEATRF"
     polyA_signal    2039..2263
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2309..2737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2751..3080
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2931..3066
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3089..3688
                     /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    3848..3969
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4018..4034)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4042..4058
                     /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(4066..4096)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4111..4132
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4420..5005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5176..6036)
                     /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(6037..6141)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6187..6566
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          6842..7858
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7952..7971
                     /label=f.pTriExUP.09.21
ORIGIN
        1 gctagccgcc accatggtat ctatcgcact tcaagccggt tatgatctcc tgggagatgg
       61 gcggcctgaa acactttggc tcgggatcgg cactttgctt atgctcattg gcacgtttta
      121 ttttctggta cgaggatggg gggtcactga caaagatgca agggagtact acgcagtgac
      181 gattcttgta agtggcatcg ccagtgctgc ttacctgtca atgtttttcg gaatcggcct
      241 cacagaggta agcgttggtg gggaaatgct cgacatttat tatgctaggt acgccgactg
      301 gttgtttacg acaccactcc tgctgctcaa cttggccttg ttggctaagg tagatagggt
      361 cactatcggc accctggtag gagtggatgc actgatgatt gtaacggggc tgattggagc
      421 cttgagtcat actgcaatcg caaggtactc atggtggctg ttctctacaa tatgtatgat
      481 tgtggtgctc tatgtgttgg ctacgtcact tcggtccgct gcacgcgaaa gaggacctga
      541 agtcgccagt acttttaata ccctgacagc tttggtgctc gtactgtgga ccgcatatcc
      601 cattctttgg attattggta cggagggagc aggtgttgtt ggactcggta tcgtgaccct
      661 cctcttcatg gttctggacg taactgcaaa ggttgggttt ggcttcatcc ttctcaggag
      721 tcgcgcaata ttgggagaca ctgagatcgg taccggcttt ccattcgacc cccattatgt
      781 ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt
      841 gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt
      901 tgcaccgacc catcgctgca ttgctccaga cctgatcggt atgggcaaat ccgacaaacc
      961 agacctgggt tatttcttcg acgaccacgt ccgcttcatg gatgccttca tcgaagccct
     1021 gggtctggaa gaggtcgtcc tggtcattca cgactggggc tccgctctgg gtttccactg
     1081 ggccaagcgc aatccagagc gcgtcaaagg tattgcattt atggagttca tccgccctat
     1141 cccgacctgg gacgaatggc cagaatttgc ccgcgagacc ttccaggcct tccgcaccac
     1201 cgacgtcggc cgcaagctga tcatcgatca gaacgttttt atcgagggta cgctgccgat
     1261 gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat taccgcgagc cgttcctgaa
     1321 tcctgttgac cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc
     1381 agcgaacatc gtcgcgctgg tcgaagaata catggactgg ctgcaccagt cccctgtccc
     1441 gaagctgctg ttctggggca ccccaggcgt tctgatccca ccggccgaag ccgctcgcct
     1501 ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc ccgggtctga atctgctgca
     1561 agaagacaac ccggacctga tcggcagcga gatcgcgcgc tggctgtcga cgctcgagat
     1621 ttccggcgag ccaaccacta agagcaggat caccagcgag ggcgagtaca tccccctgga
     1681 ccagatcgac atcaacgtgt tctgctacga gaacgaggtg caaagtcagc ctatcctgaa
     1741 cacaaaggaa atggctccac agtctaagcc tcccgaagag cttgagatgt ccagtatgcc
     1801 aagtcccgtg gctcccctcc ctgccaggac tgaaggagtg attgacatga ggagtatgtc
     1861 atctattgat agcttcatct cttgcgcaac agatttcccc gaggctactc gattctaatg
     1921 aaagcttgct accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcca
     1981 gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     2041 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2101 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2161 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2221 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2281 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2341 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2401 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2461 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2521 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2581 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2641 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2701 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2761 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2821 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2881 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2941 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3001 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3061 gaggcctagg cttttgcaaa aagctcccat gaccgagtac aagcccacgg tgcgcctcgc
     3121 cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     3181 cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga
     3241 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     3301 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     3361 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     3421 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     3481 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     3541 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     3601 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     3661 gtgcatgacc cgcaagcccg gtgcctgatt cgaaatgacc gaccaagcga cgcccaacct
     3721 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     3781 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     3841 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3901 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3961 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4021 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4081 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4141 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4201 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4261 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4321 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4381 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4441 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4501 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4561 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4621 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4681 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4741 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4801 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4861 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4921 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     4981 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5041 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5101 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5161 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5221 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5281 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5341 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5401 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5461 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5521 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5581 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5641 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5701 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5761 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5821 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5881 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5941 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6001 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6061 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6121 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     6181 ggggtcgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     6241 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     6301 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     6361 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     6421 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     6481 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     6541 acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc
     6601 ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg
     6661 cagcgatggg ggcggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
     6721 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
     6781 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
     6841 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
     6901 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
     6961 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
     7021 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
     7081 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
     7141 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     7201 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     7261 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     7321 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     7381 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     7441 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     7501 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     7561 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     7621 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     7681 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     7741 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     7801 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     7861 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcctcgagga
     7921 attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct
     7981 ggctcacaaa taccactgag atctgcatga
//
