LOCUS       Exported                7682 bp DNA     circular SYN 17-JUL-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7682)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 18, 2022 from SnapGene Viewer 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7682
                     /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             947..3202
                     /codon_start=1
                     /label=PA-wSWAPs-3x-FLAG-B-BRSBANE-60-2008
                     /translation="MDTFITRNFQTTIIQKAKNTMAEFSEDPELQPAMLFNICVHLEVC
                     YVISDMNFLDEEGKAYTALEGQGKEQNLRPQYEVIEGMPRTIAWMVQRSLAQEHGIETP
                     KYLADLFDYKTKRFIEVGITKGLADDYFWKKKEKLGNSMELMIFSYNQDYSLSNESSLD
                     EEGKGRVLSRLTELQAELSLKNLWQVLIGEEDVEKGIDFKLGQTISRLRDISVPAGFSN
                     FEGMRSYIDNIDPKGAIERNLARMSPLVSVTPKKLTWEDLRPIGPHIYDHELPEVPYNA
                     FLLMSDELGLANMTEGKSKKPKTLAKECLEKYSTLRDQTDPILIMKSEKANENFLWKLW
                     RDCVNTISNEETSNELQKTNYAKWATGDGLTYQKIMKEVAIDDETMCQEEPKIPNKCRV
                     AAWVQTEMNLLSTLTSKRALDLPEIGPDIAPVEHVGSERRKYFVNEINYCKASTVMMKY
                     VLFHTSLLNESNASMGKYKVIPITNRVVNEKGESFDMLYGLAVKGQSHLRGDTDVVTVV
                     TFEFSSTDPRVDSGKWPKYTVFRIGSLFVSGREKSVYLYCRVNGTNKIQMKWGMEARRC
                     LLQSMQQMEAIVEQESSIQGYDMTKACFKGDRVNSPKTFSIGTQEGKLVKGSFGKALRV
                     IFTKCLMHYVFGNAQLEGFSAESRRLLLLIQALKDRKGPWVFDLEGMYSGIEECISNNP
                     WVIQSVYWFNEWLGFEKEGNKVLESVDEIMDEAAADYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3107..3133)
                     /label=PA Flag Not1 Rev
     CDS             3134..3199
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(3234..3252)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3278..3502
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3548..3976
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3990..4320
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4171..4306
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4387..5181
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5355..5476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5525..5541)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5549..5565
                     /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(5573..5603)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5618..5639
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5927..6515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6686..7546)
                     /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(7547..7651)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcaccatgg atacttttat
      961 tacaagaaac ttccagacta caataataca aaaggccaaa aacacaatgg cagaatttag
     1021 tgaagatcct gaattgcaac cagcaatgct attcaatatc tgcgtccatc tagaggtttg
     1081 ctatgtaata agtgacatga attttcttga cgaagaagga aaagcatata cagcattaga
     1141 aggacaaggg aaagaacaaa acttgagacc acaatatgaa gtaattgagg gaatgccaag
     1201 aaccatagca tggatggtcc agagatcctt agctcaagag catggaatag agactcccaa
     1261 gtatctggct gatttgtttg attataaaac caaaagattt atagaagttg gaataacaaa
     1321 gggattggct gatgattact tttggaaaaa gaaagaaaag ttgggaaata gcatggaact
     1381 gatgatattc agctacaatc aagactactc gttaagtaat gaatcctcat tggatgagga
     1441 agggaaaggg agagtgctaa gcagactcac agaacttcag gctgaattaa gtctgaaaaa
     1501 tttatggcaa gttctcatag gagaagaaga tgttgaaaag ggaattgatt ttaaacttgg
     1561 acaaacaata tctagactaa gggatatatc tgttccagct ggtttctcca attttgaagg
     1621 aatgaggagc tacatagaca atatagaccc aaaaggagca atagagagaa atctagcaag
     1681 gatgtctccc ttagtatcag tcacacctaa aaagttaaca tgggaggacc taagaccaat
     1741 agggcctcac atttacgacc atgagctacc agaagttcca tataatgcct ttcttctaat
     1801 gtctgatgaa ctgggattgg ccaatatgac tgagggaaag tccaaaaaac cgaagacatt
     1861 agccaaagaa tgtctagaaa agtactcaac actacgggat caaactgacc caatattaat
     1921 aatgaaaagc gaaaaagcta acgaaaattt cctatggaag ctttggagag actgtgtaaa
     1981 tacaataagt aatgaggaaa cgagtaacga gttacagaaa accaattatg ccaaatgggc
     2041 cacaggggat ggattaacat accagaaaat aatgaaagaa gtagcaatag atgacgaaac
     2101 aatgtgccaa gaagagccta aaatccctaa caaatgtaga gtggctgctt gggttcaaac
     2161 agagatgaat ctattgagca ctctgacaag taaaagagct ctggacctac cagaaatagg
     2221 gccagacata gcacccgtgg agcatgtagg aagtgaaaga aggaaatact ttgttaatga
     2281 aatcaactac tgtaaggcct ctacagttat gatgaagtat gtgctttttc acacttcatt
     2341 gttgaatgaa agcaatgcca gcatgggaaa atacaaagta ataccaataa ccaatagagt
     2401 agtaaatgaa aaaggagaaa gtttcgacat gctttacggt ctggcggtta aaggacaatc
     2461 tcatctgagg ggagatactg atgttgtaac agttgtaact ttcgaattta gtagtacaga
     2521 tccaagagtg gactcaggaa agtggccaaa atatactgtg tttaggattg gctccctatt
     2581 tgtgagtggg agggaaaaat ctgtgtactt gtattgccga gtgaatggca caaataagat
     2641 ccaaatgaaa tggggaatgg aagctagaag atgtttgctt caatcaatgc aacaaatgga
     2701 ggcaattgtt gaacaggaat catcaataca aggatatgac atgaccaaag cctgtttcaa
     2761 gggagacaga gtaaatagcc ccaaaacttt cagtattgga actcaagaag gaaaactagt
     2821 aaaaggatcc tttggaaaag cactaagagt aatatttact aaatgcttga tgcactatgt
     2881 atttggaaat gcccaattgg aggggtttag tgccgagtct aggagacttc tactgttgat
     2941 tcaagcatta aaggacagaa agggcccttg ggtgttcgac ttagagggaa tgtattctgg
     3001 aatagaagaa tgtattagca acaacccttg ggtaatacag agtgtatact ggttcaatga
     3061 atggttgggc tttgagaagg agggcaacaa ggttctcgaa agtgtagacg agattatgga
     3121 cgaagcggcc gccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     3181 caaggatgac gatgacaagt aggccgctcg agcatgcatc tagagggccc tattctatag
     3241 tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     3301 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3361 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3421 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3481 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3541 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3601 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3661 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     3721 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3781 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3841 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3901 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3961 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4021 gctccccagg caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4081 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4141 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4201 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4261 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4321 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4381 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4441 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4501 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4561 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4621 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4681 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4741 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4801 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4861 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4921 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4981 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5041 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5101 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5161 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5221 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5281 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5341 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5401 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5461 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5521 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5581 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5641 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5701 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5761 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5821 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5881 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5941 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6001 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6061 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6121 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6181 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6241 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6301 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6361 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6421 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6481 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6541 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6601 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6661 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6721 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6781 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6841 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6901 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6961 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7021 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7081 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7141 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7201 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7261 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7321 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7381 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7441 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7501 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7561 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7621 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7681 tc
//
