LOCUS       Exported                7343 bp ds-DNA     circular SYN 17-FEB-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7343)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Feb 17, 2021 from SnapGene Viewer 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7343
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 143 .. 148 / #ffffff / -35
                        2: 149 .. 166 / #ffffff
                        3: 167 .. 173 / #ffffff / -10"
     protein_bind    181..197
                     /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)."
                     /note="color: #31849b"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     misc_feature    248..378
                     /label=ITR
                     /note="color: #800000"
     misc_feature    476..671
                     /label=RHO
                     /note="color: #00ff00; direction: RIGHT"
     regulatory      696..705
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             698..853
                     /codon_start=1
                     /label=RmaIntC
                     /note="color: #0000ff"
                     /translation="MAAACPELRQLAQSDVYWDPIVSIEPDGVEEVFDLTVPGPHNFVA
                     NDIIAHN"
     CDS             854..2851
                     /codon_start=1
                     /label=dCas9C
                     /note="color: #993366"
                     /translation="SGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPEN
                     IVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQ
                     NGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVV
                     KKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQIL
                     DSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVG
                     TALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLA
                     NGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILP
                     KRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMER
                     SSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPS
                     KYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDK
                     VLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLI
                     HQSITGLYETRIDLSQLGGDSRADPKKKRKV"
     CDS             1302..1313
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 1313"
                     /translation="IEGR"
     CDS             2831..2851
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     protein_bind    2888..2912
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
                     /note="color: #31849b; direction: RIGHT"
     misc_feature    2942..4510
                     /label=VPR
                     /note="color: #00ffff; direction: RIGHT"
     misc_feature    4515..4562
                     /label=pA terminator
                     /note="color: #a6acb3"
     misc_feature    4649..4779
                     /label=ITR
                     /note="color: #800000"
     primer_bind     complement(4798..4814)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     rep_origin      4956..5411
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        5438..5542
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             5543..6403
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5543 .. 5611 / #ccffcc / signal sequence
                        2: 5612 .. 6403 / #ccffcc
                     Cleavage site after base 5611"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVVMVTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6574..7162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc
      301 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
      361 catcactagg ggttccttgt agttaatgat taacccgcca tgctacttat ctacgtagcc
      421 atgctctagg aagatcggaa ttcgccctta agggcgcgcc gtttaaatag ctagccctct
      481 cctccctgac ctcaggcttc ctcctagtgt caccttggcc cctcttagaa gccaattagg
      541 ccctcagttt ctgcagcggg gattaatatg attatgaaca cccccaatct cccagatgct
      601 gattcagcca ggagcttagg agggggaggt cactttataa gggtctgggg gggtcagaac
      661 ccagagtcat cggtaccacc ggtcgactag aggatccatg gcggcggcgt gcccggaact
      721 gcgtcagctg gcgcagagcg atgtgtattg ggatccgatt gtgagcattg aaccggatgg
      781 cgtggaagaa gtgtttgatc tgaccgtgcc gggcccgcat aactttgtgg cgaacgatat
      841 tattgcgcat aactctggcc agggggacag tcttcacgag cacatcgcta atcttgcagg
      901 tagcccagct atcaaaaagg gaatactgca gaccgttaag gtcgtggatg aactcgtcaa
      961 agtaatggga aggcataagc ccgagaatat cgttatcgag atggcccgag agaaccaaac
     1021 tacccagaag ggacagaaga acagtaggga aaggatgaag aggattgaag agggtataaa
     1081 agaactgggg tcccaaatcc ttaaggaaca cccagttgaa aacacccagc ttcagaatga
     1141 gaagctctac ctgtactacc tgcagaacgg cagggacatg tacgtggatc aggaactgga
     1201 catcaatcgg ctctccgact acgacgtgga tgctatcgtg ccccagtctt ttctcaaaga
     1261 tgattctatt gataataaag tgttgacaag atccgataaa aatagaggga agagtgataa
     1321 cgtcccctca gaagaagttg tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc
     1381 caaactgatc acacaacgga agttcgataa tctgactaag gctgaacgag gtggcctgtc
     1441 tgagttggat aaagccggct tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa
     1501 gcacgtggcc caaattctcg attcacgcat gaacaccaag tacgatgaaa atgacaaact
     1561 gattcgagag gtgaaagtta ttactctgaa gtctaagctg gtctcagatt tcagaaagga
     1621 ctttcagttt tataaggtga gagagatcaa caattaccac catgcgcatg atgcctacct
     1681 gaatgcagtg gtaggcactg cacttatcaa aaaatatccc aagcttgaat ctgaatttgt
     1741 ttacggagac tataaagtgt acgatgttag gaaaatgatc gcaaagtctg agcaggaaat
     1801 aggcaaggcc accgctaagt acttctttta cagcaatatt atgaattttt tcaagaccga
     1861 gattacactg gccaatggag agattcggaa gcgaccactt atcgaaacaa acggagaaac
     1921 aggagaaatc gtgtgggaca agggtaggga tttcgcgaca gtccggaagg tcctgtccat
     1981 gccgcaggtg aacatcgtta aaaagaccga agtacagacc ggaggcttct ccaaggaaag
     2041 tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa
     2101 gaaatacggc ggattcgatt ctcctacagt cgcttacagt gtactggttg tggccaaagt
     2161 ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat
     2221 ggagcgatca agcttcgaaa aaaaccccat cgactttctc gaggcgaaag gatataaaga
     2281 ggtcaaaaaa gacctcatca ttaagcttcc caagtactct ctctttgagc ttgaaaacgg
     2341 ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc
     2401 ctctaaatac gttaatttct tgtatctggc cagccactat gaaaagctca aagggtctcc
     2461 cgaagataat gagcagaagc agctgttcgt ggaacaacac aaacactacc ttgatgagat
     2521 catcgagcaa ataagcgaat tctccaaaag agtgatcctc gccgacgcta acctcgataa
     2581 ggtgctttct gcttacaata agcacaggga taagcccatc agggagcagg cagaaaacat
     2641 tatccacttg tttactctga ccaacttggg cgcgcctgca gccttcaagt acttcgacac
     2701 caccatagac agaaagcggt acacctctac aaaggaggtc ctggacgcca cactgattca
     2761 tcagtcaatt acggggctct atgaaacaag aatcgacctc tctcagctcg gtggagacag
     2821 cagggctgac cccaagaaga agaggaaggt gtcgccaggg atccgtcgac ttgacgcgtt
     2881 gatatcaaca agtttgtaca aaaaagcagg ctacaaagag gccagcggtt ccggacgggc
     2941 tgacgcattg gacgattttg atctggatat gctgggaagt gacgccctcg atgattttga
     3001 ccttgacatg cttggttcgg atgcccttga tgactttgac ctcgacatgc tcggcagtga
     3061 cgcccttgat gatttcgacc tggacatgct gattaactct agaagttccg gatctccgaa
     3121 aaagaaacgc aaagttggta gccagtacct gcccgacacc gacgaccggc accggatcga
     3181 ggaaaagcgg aagcggacct acgagacatt caagagcatc atgaagaagt cccccttcag
     3241 cggccccacc gaccctagac ctccacctag aagaatcgcc gtgcccagca gatccagcgc
     3301 cagcgtgcca aaacctgccc cccagcctta ccccttcacc agcagcctga gcaccatcaa
     3361 ctacgacgag ttccctacca tggtgttccc cagcggccag atctctcagg cctctgctct
     3421 ggctccagcc cctcctcagg tgctgcctca ggctcctgct cctgcaccag ctccagccat
     3481 ggtgtctgca ctggctcagg caccagcacc cgtgcctgtg ctggctcctg gacctccaca
     3541 ggctgtggct ccaccagccc ctaaacctac acaggccggc gagggcacac tgtctgaagc
     3601 tctgctgcag ctgcagttcg acgacgagga tctgggagcc ctgctgggaa acagcaccga
     3661 tcctgccgtg ttcaccgacc tggccagcgt ggacaacagc gagttccagc agctgctgaa
     3721 ccagggcatc cctgtggccc ctcacaccac cgagcccatg ctgatggaat accccgaggc
     3781 catcacccgg ctcgtgacag gcgctcagag gcctcctgat ccagctcctg cccctctggg
     3841 agcaccaggc ctgcctaatg gactgctgtc tggcgacgag gacttcagct ctatcgccga
     3901 tatggatttc tcagccttgc tgggctctgg cagcggcagc cgggattcca gggaagggat
     3961 gtttttgccg aagcctgagg ccggctccgc tattagtgac gtgtttgagg gccgcgaggt
     4021 gtgccagcca aaacgaatcc ggccatttca tcctccagga agtccatggg ccaaccgccc
     4081 actccccgcc agcctcgcac caacaccaac cggtccagta catgagccag tcgggtcact
     4141 gaccccggca ccagtccctc agccactgga tccagcgccc gcagtgactc ccgaggccag
     4201 tcacctgttg gaggatcccg atgaagagac gagccaggct gtcaaagccc ttcgggagat
     4261 ggccgatact gtgattcccc agaaggaaga ggctgcaatc tgtggccaaa tggacctttc
     4321 ccatccgccc ccaaggggcc atctggatga gctgacaacc acacttgagt ccatgaccga
     4381 ggatctgaac ctggactcac ccctgacccc ggaattgaac gagattctgg ataccttcct
     4441 gaacgacgag tgcctcttgc atgccatgca tatcagcaca ggactgtcca tcttcgacac
     4501 atctctgttt tgacaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     4561 gtctcgagtt aagggcgaat tcccgattag gatcttccta gagcatggct acgtagataa
     4621 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc
     4681 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     4741 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc cttaattaac ctaattcact
     4801 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     4861 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     4921 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     4981 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5041 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5101 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5161 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccccgataga cggtttttcg
     5221 ccctttgacg ctggagttca cgttcctcaa tagtggactc ttgttccaaa ctggaacaac
     5281 actcaaccct atctcggtct attcttttga tttataaggg atttttccga tttcggccta
     5341 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     5401 gtttataatt tcaggtggca tctttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5461 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5521 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5581 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5641 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca atagtggtaa
     5701 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5761 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5821 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5881 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5941 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6001 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6061 cgacgagcgt gacaccacga tgcctgtagt aatggtaaca acgttgcgca aactattaac
     6121 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6181 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6241 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6301 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6361 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6421 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6481 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6541 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6601 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6661 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6721 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6781 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6841 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6901 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6961 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7021 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7081 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7141 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7201 ttgctgcggt tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7261 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     7321 gtcagtgagc gaggaagcgg aag
//
