LOCUS       Exported                7572 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 7572)
  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..7572
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1397..1552
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    14..1480
                     /label=Ace2_D92N_E199V-mRuby3
     CDS             14..694
                     /codon_start=1
                     /label=Ace2_D92N_E199V
                     /translation="MADVETETGMIAQWIVFAIMAAAAIAFGVAVHFRPSELKSAYYIN
                     IAICTIAATAYYAMAVNYQDLTMNGERQVVYARYIDWVLTTPLLLLNLIVMTKMGGVMI
                     SWVIGADIFMIVFGILGAFEDEHKFKWVYFIAGCVMQAVLTYGMYNATWKDDLKKSPEY
                     HSSYVSLLVFLSILWVFYPVVWAFGSGSGVLSVDNVAILMGILDVLAKPLFGMGCLIAH
                     ETIFK"
     CDS             695..706
                     /codon_start=1
                     /label=linker
                     /translation="WRSG"
     CDS             707..1396
                     /codon_start=1
                     /label=mRuby3 (minus 7 aa in Nt)
                     /note="mRuby3"
                     /translation="LIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMRIKVIEG
                     GPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGVVTVTQD
                     TSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDIALKVDG
                     GGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVAKYSNLG
                     GGMDELYK"
     CDS             1397..1456
                     /codon_start=1
                     /label=TS from Kir2.1
                     /translation="KSRITSEGEYIPLDQIDINV"
     CDS             1457..1477
                     /codon_start=1
                     /label=ER export sequence from Kir2.1
                     /translation="FCYENEV"
     primer_bind     complement(1595..1612)
                     /label=r-FAR
     polyA_signal    1601..1825
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1871..2299
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2313..2642
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     2488..2500
                     /label=Primer 1
     rep_origin      2493..2628
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2651..3250
                     /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    3410..3531
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3580..3596)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3604..3620
                     /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(3628..3658)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3673..3694
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3982..4567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4738..5598)
                     /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(5599..5703)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5749..6128
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          6404..7420
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7514..7533
                     /label=f.pTriExUP.09.21
ORIGIN
        1 gctagccgcc accatggctg acgtggaaac cgagaccggc atgattgcac agtggattgt
       61 ctttgctatt atggctgctg ctgctattgc ttttggagtg gctgtgcact ttcggccttc
      121 agagctgaag agcgcatact atatcaacat tgccatctgc actatcgccg ctaccgctta
      181 ctatgcaatg gccgtgaact accaggacct gacaatgaat ggtgaaaggc aggtggtcta
      241 cgcaagatat attgactggg tgctgaccac accactgctc ctgctcaacc tcatcgtcat
      301 gaccaagatg ggcggagtga tgatttcttg ggtcatcggc gcagacattt tcatgatcgt
      361 gtttggtatt ctgggcgcct tcgaggatga acacaagttc aaatgggtgt actttatcgc
      421 tggatgtgtg atgcaggcag tcctgacata cgggatgtat aacgccactt ggaaagacga
      481 tctgaagaaa agccccgagt accatagctc ctatgtcagt ctgctcgtct tcctgtcaat
      541 cctctgggtg ttttatcctg tcgtgtgggc tttcgggtct ggtagtggcg tgctgtccgt
      601 cgacaatgtg gccattctca tgggaatcct ggatgtgctc gctaagccac tgtttggaat
      661 ggggtgcctc attgcccatg agactatctt caagtggagg tccggactga tcaaggaaaa
      721 tatgcgtatg aaggtggtca tggaaggttc ggtcaacggc caccaattca aatgcacagg
      781 tgaaggagaa ggcaggccgt acgagggagt gcaaaccatg aggatcaaag tcatcgaggg
      841 aggacccctg ccatttgcct ttgacattct tgccacgtcg ttcatgtatg gcagccgtac
      901 ttttatcaag tacccggcag acattcctga tttctttaaa cagtcctttc ctgagggttt
      961 tacttgggaa agagttacga gatacgaaga tggtggagtc gtcaccgtca cgcaggacac
     1021 cagccttgag gatggcgaac tcgtttacaa cgtcaaagtc agaggggtaa actttccctc
     1081 caatggtccc gtgatgcaga agaagaccaa gggttgggag cctaatacag agatgatgta
     1141 tccagcagat ggtggtctga ggggatacac tgatattgca ctgaaagttg atggtggtgg
     1201 ccatctgcat tgcaatttcg taacaactta caggtcaaaa aagaccgtcg ggaacatcaa
     1261 gatgcccggt gtccatgccg ttgatcaccg cctggaaagg atagaggaaa gtgacaatga
     1321 aacgtacgta gtacaacgcg aagtcgcagt tgccaagtac tccaatcttg gtggtggtat
     1381 ggatgaactt tataagaaga gcaggatcac cagcgagggc gagtacatcc ccctggacca
     1441 gatcgatatc aacgtgttct gctacgagaa cgaggtgtaa tgaaagcttg ctaccgagct
     1501 cggatccact agtccagtgt ggtggaattc tgcagaaatc cagcacagtg gcggccgctc
     1561 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     1621 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     1681 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     1741 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     1801 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     1861 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1921 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1981 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2041 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2101 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2161 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2221 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2281 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2341 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2401 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2461 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2521 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2581 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     2641 aaaagctccc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     2701 ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     2761 cgacccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     2821 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     2881 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     2941 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     3001 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     3061 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     3121 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     3181 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     3241 cggtgcctga ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3301 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3361 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     3421 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3481 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3541 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3601 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3661 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3721 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3781 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3841 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3901 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3961 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4021 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4081 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4141 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4201 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4261 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4321 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4381 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4441 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4501 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4561 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4621 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4681 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4741 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4801 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4861 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4921 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4981 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5041 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5101 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5161 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5221 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5281 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5341 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5401 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5461 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5521 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5581 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5641 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5701 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcggggtcga cattgattat
     5761 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     5821 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     5881 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     5941 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     6001 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     6061 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     6121 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     6181 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
     6241 gggggggcgc gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag
     6301 aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg
     6361 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg
     6421 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
     6481 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg
     6541 cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg
     6601 ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg
     6661 gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg
     6721 gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg
     6781 cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
     6841 agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc
     6901 tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt
     6961 gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg
     7021 ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc
     7081 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
     7141 ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg
     7201 ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc
     7261 gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct
     7321 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc
     7381 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     7441 tattgtgctg tctcatcatt ttggcaaaga attcctcgag gaattcactc ctcaggtgca
     7501 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     7561 agatctgcat ga
//
