LOCUS       62056_1745        8113 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8113)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             929..949
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             953..3610
                     /codon_start=1
                     /label=NgAgo
                     /note="Argonaute protein from Natronobacterium gregoryi
                     (Gao et al., 2016)"
                     /translation="TVIDLDSTTTADELTSGHTYDISVTLTGVYDNTDEQHPRMSLAFE
                     QDNGERRYITLWKNTTPKDVFTYDYATGSTYIFTNIDYEVKDGYENLTATYQTTVENAT
                     AQEVGTTDEDETFAGGEPLDHHLDDALNETPDDAETESDSGHVMTSFASRDQLPEWTLH
                     TYTLTATDGAKTDTEYARRTLAYTVRQELYTDHDAAPVATDGLMLLTPEPLGETPLDLD
                     CGVRVEADETRTLDYTTAKDRLLARELVEEGLKRSLWDDYLVRGIDEVLSKEPVLTCDE
                     FDLHERYDLSVEVGHSGRAYLHINFRHRFVPKLTLADIDDDNIYPGLRVKTTYRPRRGH
                     IVWGLRDECATDSLNTLGNQSVVAYHRNNQTPINTDLLDAIEAADRRVVETRRQGHGDD
                     AVSFPQELLAVEPNTHQIKQFASDGFHQQARSKTRLSASRCSEKAQAFAERLDPVRLNG
                     STVEFSSEFFTGNNEQQLRLLYENGESVLTFRDGARGAHPDETFSKGIVNPPESFEVAV
                     VLPEQQADTCKAQWDTMADLLNQAGAPPTRSETVQYDAFSSPESISLNVAGAIDPSEVD
                     AAFVVLPPDQEGFADLASPTETYDELKKALANMGIYSQMAYFDRFRDAKIFYTRNVALG
                     LLAAAGGVAFTTEHAMPGDADMFIGIDVSRSYPEDGASGQINIAATATAVYKDGTILGH
                     SSTRPQLGEKLQSTDVRDIMKNAILGYQQVTGESPTHIVIHRDGFMNEDLDPATEFLNE
                     QGVEYDIVEIRKQPQTRLLAVSDVQYDTPVKSIAAINQNEPRATVATFGAPEYLATRDG
                     GGLPRPIQIERVAGETDIETLTRQVYLLSQSHIQVHNSTARLPITTAYADQASTHATKG
                     YLVQTGAFESNVGFL"
     polyA_signal    3713..3937
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3983..4411
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4425..4754
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4821..5612
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5789..5922
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5959..5975)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5983..5999)
                     /label=lac operator
                     /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(6007..6037)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6052..6073)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6361..6946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7120..7977)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7978..8082)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatggtgcc aaaaaagaag agaaaggtag ccacagtgat
      961 tgacctcgat tcgaccacca ccgcagacga actgacatcg ggacacacgt acgacatctc
     1021 agtcacgctc accggtgtct acgataacac cgacgagcag catcctcgca tgtctctcgc
     1081 attcgagcag gacaacggcg agcggcgtta cattaccctg tggaagaaca cgacacccaa
     1141 ggatgtcttt acatacgact acgccacggg ctcgacgtac atcttcacta acatcgacta
     1201 cgaagtgaag gacggctacg agaatctgac tgcaacatac cagacgaccg tcgagaacgc
     1261 taccgctcag gaagtcggga cgactgacga ggacgaaacg ttcgcgggcg gcgagccgct
     1321 cgaccatcac ttggacgacg cgctcaatga gacgccagac gacgcggaga cagagagcga
     1381 ctcaggccat gtgatgacct cgttcgcctc ccgcgaccaa ctccctgagt ggacgctgca
     1441 tacgtatacg ctaacagcca cagacggcgc aaagacggac acggagtacg cgcgacgaac
     1501 cctcgcatac acggtacggc aggaactcta taccgaccat gatgcggctc cggttgcaac
     1561 tgacgggcta atgcttctca cgccagagcc gctcggcgag accccgcttg acctcgattg
     1621 cggtgtccgg gtcgaggcgg acgagactcg gacactcgat tacaccacgg ccaaagaccg
     1681 gttactcgcc cgcgaactcg tcgaagaggg gctcaaacgc tccctctggg atgactacct
     1741 cgttcgcggc atcgatgaag tcctctcaaa ggagcctgtg ctgacttgcg atgagttcga
     1801 cctacatgag cggtatgacc tctctgtcga agtcggtcac agtgggcggg cgtaccttca
     1861 catcaacttc cgccaccggt tcgtaccgaa gctgacgctc gcagacatcg atgatgacaa
     1921 catctatcct gggctccggg tgaagacgac gtatcgcccc cggcgaggac atatcgtctg
     1981 gggtctgcgg gacgagtgcg ccaccgactc gctcaacacg ctgggaaacc agtccgtcgt
     2041 tgcataccac cgcaacaatc agacacctat taacactgac ctcctcgacg ctatcgaggc
     2101 cgctgaccgg cgagtcgtcg aaacccgacg tcaagggcac ggcgatgatg ctgtctcatt
     2161 cccccaagaa ctgcttgcgg tcgaaccgaa tacgcaccaa attaagcagt tcgcctccga
     2221 cggattccac caacaggccc gctcaaagac gcgtctctcg gcctcccgct gcagcgagaa
     2281 agcgcaagcg ttcgccgagc ggcttgaccc ggtgcgtctc aatgggtcca cggtagagtt
     2341 ctcctcggag tttttcaccg ggaacaacga gcagcaactg cgcctcctct acgagaacgg
     2401 tgagtcggtt ctgacgttcc gcgacggggc gcgtggtgcg caccccgacg agacattctc
     2461 gaaaggtatc gtcaatccac cagagtcgtt cgaggtggcc gtagtactgc ccgagcagca
     2521 ggcagatacc tgcaaagcgc agtgggacac gatggctgac ctcctcaacc aagctggcgc
     2581 gccaccgaca cggagcgaga ccgtccaata tgatgcgttc tcctcgccag agagcatcag
     2641 cctcaatgtg gctggagcca tcgaccctag cgaggtagac gcggcattcg tcgtactgcc
     2701 gccggaccaa gaaggattcg cagacctcgc cagtccgaca gagacgtacg acgagctgaa
     2761 gaaggcgctt gccaacatgg gcatttacag ccagatggcg tacttcgacc ggttccgcga
     2821 cgcgaaaata ttctatactc gtaacgtggc actcgggctg ctggcagccg ctggcggcgt
     2881 cgcattcaca accgaacatg cgatgcctgg ggacgcagat atgttcattg ggattgatgt
     2941 ctctcggagc taccccgagg acggtgccag cggccagata aacattgccg cgacggcgac
     3001 cgccgtctac aaggatggaa ctatcctcgg ccactcgtcc acccgaccgc agctcgggga
     3061 gaaactacag tcgacggatg ttcgtgacat tatgaagaat gccatcctcg gctaccagca
     3121 ggtgaccggt gagtcgccga cccatatcgt catccaccgt gatggcttca tgaacgaaga
     3181 cctcgacccc gccacggaat tcctcaacga acaaggcgtc gagtacgaca tcgtcgaaat
     3241 ccgcaagcag ccccagacac gcctgctggc agtctccgat gtgcagtacg atacgcctgt
     3301 gaagagcatc gccgctatca accagaacga gccacgggca acggtcgcca ccttcggcgc
     3361 acccgaatac ttagcgacac gcgatggagg cggccttccc cgcccaatcc aaattgaacg
     3421 agtcgccggc gaaaccgaca tcgagacgct cactcgccaa gtctatctgc tctcccagtc
     3481 gcatatccag gtccataact cgactgcgcg cctacccatc accaccgcat acgccgacca
     3541 ggcaagtact cacgcgacca agggttacct cgtccagacc ggagcgttcg agtctaatgt
     3601 cggattcctc tgaggatcca ctagtccagt gtggtggaat tctgcagata tccagcacag
     3661 tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     3721 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3781 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3841 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3901 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3961 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4021 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4081 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4141 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4201 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4261 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4321 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4381 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4441 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4501 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4561 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4621 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4681 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4741 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4801 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4861 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4921 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4981 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5041 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5101 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5161 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5221 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5281 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5341 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5401 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5461 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5521 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5581 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5641 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5701 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5761 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5821 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5881 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5941 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6001 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6061 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6121 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6181 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6241 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6301 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6361 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6421 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6481 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6541 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6601 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6661 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6721 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6781 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6841 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     6901 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6961 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7021 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7081 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7141 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7201 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7261 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7321 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7381 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7441 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7501 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7561 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7621 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7681 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7741 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7801 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7861 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7921 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7981 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8041 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8101 gccacctgac gtc
//
