LOCUS       Exported                7351 bp ds-DNA     circular SYN 24-JUN-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7351)
  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..7351
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      8..17
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             14..1465
                     /label=Ace2_D92N_E199V-mNeon
     CDS             14..706
                     /codon_start=1
                     /label=Ace2_D92N_E199V
                     /translation="MADVETETGMIAQWIVFAIMAAAAIAFGVAVHFRPSELKSAYYIN
                     IAICTIAATAYYAMAVNYQDLTMNGERQVVYARYIDWVLTTPLLLLNLIVMTKMGGVMI
                     SWVIGADIFMIVFGILGAFEDEHKFKWVYFIAGCVMQAVLTYGMYNATWKDDLKKSPEY
                     HSSYVSLLVFLSILWVFYPVVWAFGSGSGVLSVDNVAILMGILDVLAKPLFGMGCLIAH
                     ETIFKKMLR"
     CDS             707..1381
                     /codon_start=1
                     /label=mNeonGreen
                     /translation="SLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEELNLKSTKGDL
                     QFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDGASLTVNYRYT
                     YEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIISTFKWSYTTGNG
                     KRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQKAFTDVMGMDE
                     LYK"
     regulatory      929..938
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1382..1441
                     /codon_start=1
                     /label=TS from Kir2.1
                     /translation="KSRITSEGEYIPLDQIDINV"
     CDS             1442..1465
                     /codon_start=1
                     /label=ER export sequence from Kir2.1
                     /translation="FCYENEV"
     intron          1531..1663
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    1672..2260
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2143..2154)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2293..2404
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2696..2751
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3110..3126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3134..3150
                     /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(3158..3188)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3203..3224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3282..3478
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3329..3464
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3484..3618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3857..4445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4616..5476)
                     /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(5477..5581)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5611..5990
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5993..6268
                     /label=chicken beta-actin promoter
     intron          6269..7277
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
ORIGIN
        1 cggtgccgcc 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 caagaagatg ctgaggtctc tcccagcgac
      721 acatgagtta cacatctttg gctccatcaa cggtgtggac tttgacatgg tgggtcaggg
      781 caccggcaat ccaaatgatg gttatgagga gttaaacctg aagtccacca agggtgacct
      841 ccagttctcc ccctggattc tggtccctca tatcgggtat ggcttccatc agtacctgcc
      901 ctaccctgac gggatgtcgc ctttccaggc cgccatggta gatggctccg gataccaagt
      961 ccatcgcaca atgcagtttg aagatggtgc ctcccttact gttaactacc gctacaccta
     1021 cgagggaagc cacatcaaag gagaggccca ggtgaagggg actggtttcc ctgctgacgg
     1081 tcctgtgatg accaactcgc tgaccgctgc ggactggtgc aggtcgaaga agacttaccc
     1141 caacgacaaa accatcatca gtacctttaa gtggagttac accactggaa atggcaagcg
     1201 ctacaggagc actgcgcgga ccacctacac ctttgccaag ccaatggcgg ctaactatct
     1261 gaagaaccag ccgatgtacg tgttccgtaa gacggagctc aagcactcca agaccgagct
     1321 caacttcaag gagtggcaaa aggcctttac cgatgtgatg ggcatggacg agctgtacaa
     1381 gaagagcagg atcaccagcg agggcgagta catccccctg gaccagatcg acatcaacgt
     1441 gttctgctac gagaacgagg tgtaagaatt cgatatcggt accgatgaat agctcgaggt
     1501 accgatgaat agctaaggtc gaggccgcag gtaagtatca aggttacaag acaggtttaa
     1561 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
     1621 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcga caatcaacct
     1681 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     1741 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     1801 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     1861 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     1921 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     1981 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2041 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     2101 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2161 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     2221 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattcg agctcggtac
     2281 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2341 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2401 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2461 agggggagga ttgggaagac aatagcaggc cagcttttgt tccctttagg gttaattgcg
     2521 cgcttggcgc taatcatggt catagctgtt tccactcctc aggtgcaggc tgcctatcag
     2581 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     2641 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     2701 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     2761 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     2821 acatatgcca tatgctggct gccatgaaca aaggtggcta taaagaggtc atcagtatat
     2881 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat
     2941 ttttttttat attttgtttg tgttattttt ttctttaaca tccctaaaat tttccttaca
     3001 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct
     3061 tctcttatga agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt
     3121 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3181 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3241 tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc
     3301 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3361 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3421 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3481 gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3541 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3601 tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3661 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3721 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3781 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3841 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3901 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3961 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4021 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     4081 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4141 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4201 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4261 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     4321 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4381 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4441 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4501 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4561 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4621 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4681 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4741 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4801 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4861 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4921 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4981 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5041 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5101 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5161 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5221 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5281 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5341 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5401 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5461 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5521 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5581 gcacatttcc ccgaaaagtg ccacctggtc gacattgatt attgactagt tattaatagt
     5641 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     5701 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     5761 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggactatt
     5821 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     5881 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     5941 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ggtcgaggtg
     6001 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
     6061 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggcgcgcgcc
     6121 aggcggggcg gggcgggggc gagggcgggg cggggcgagg cggagaggtg cggcggcagc
     6181 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
     6241 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg ttgccttcgc cccgtgcccc
     6301 gctccgcgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
     6361 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct
     6421 cgtttctttt ctgtggctgc gtgaaagcct taaagggctc cgggagggcc ctttgtgcgg
     6481 gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggcc
     6541 cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcgt
     6601 gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggc tgcgagggga
     6661 acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcggcgg
     6721 tcgggctgta acccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg
     6781 ggtgcggggc tccgtgcggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg
     6841 caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggagggctc gggggagggg
     6901 cgcggcggcc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt
     6961 tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgg cggagccgaa
     7021 atctgggagg cgccgccgca ccccctctag cgggcgcggg cgaagcggtg cggcgccggc
     7081 aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc ccttctccat
     7141 ctccagcctc ggggctgccg cagggggacg gctgccttcg ggggggacgg ggcagggcgg
     7201 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
     7261 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
     7321 gcaaagaatt cggtaccgcg ggcccgggaa c
//
