LOCUS       Exported                5757 bp ds-DNA     circular SYN 26-MAR-2020
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5757)
  TITLE     Baird et al., 2000 (R95K mutation)
REFERENCE   2  (bases 1 to 5757)
  TITLE     Bintu et al., 2016
REFERENCE   3  (bases 1 to 5757)
  TITLE     Bintu et al., 2016
REFERENCE   4  (bases 1 to 5757)
  TITLE     Hermann, et al. 2014
REFERENCE   5  (bases 1 to 5757)
  TITLE     Xu et al., 2009, Donahue et al., 2020
REFERENCE   6  (bases 1 to 5757)
  TITLE     Hansen et al., 2014
REFERENCE   7  (bases 1 to 5757)
  TITLE     Xu et al., 2009, Donahue et al., 2020
REFERENCE   8  (bases 1 to 5757)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   9  (bases 1 to 5757)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   10  (bases 1 to 5757)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   11  (bases 1 to 5757)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   12  (bases 1 to 5757)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   13  (bases 1 to 5757)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   14  (bases 1 to 5757)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   15  (bases 1 to 5757)
  AUTHORS   Joe Muldoon
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Sep 28, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: ""Unpublished Work""
            SGRef: number: 2; type: ""Unpublished Work""
            SGRef: number: 3; type: ""Unpublished Work""
            SGRef: number: 4; type: ""Unpublished Work""
            SGRef: number: 5; type: ""Unpublished Work""
            SGRef: number: 6; type: ""Unpublished Work""
            SGRef: number: 7; type: ""Unpublished Work""
            SGRef: number: 8; type: ""Unpublished Work""
            SGRef: number: 9; type: ""Unpublished Work""
            SGRef: number: 10; type: ""Unpublished Work""
            SGRef: number: 11; type: ""Unpublished Work""
            SGRef: number: 12; type: ""Unpublished Work""
            SGRef: number: 13; type: ""Unpublished Work""
            SGRef: number: 14; type: ""Unpublished Work""
FEATURES             Location/Qualifiers
     source          1..5757
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    31..55
                     /label=TUPV2 Barcode F
                     /note="TUPV barcode"
                     /citation="[5]"
     misc_feature    68..76
                     /label=ZF6 BS
                     /note="Binding site for ZF150-4"
                     /citation="[8]"
     misc_feature    83..91
                     /label=ZF6 BS
                     /note="Binding site for ZF150-4"
                     /citation="[9]"
     misc_feature    98..106
                     /label=ZF6 BS
                     /note="Binding site for ZF150-4"
                     /citation="[10]"
     misc_feature    113..121
                     /label=ZF6 BS
                     /note="Binding site for ZF150-4"
                     /citation="[11]"
     misc_feature    128..136
                     /label=ZF6 BS
                     /note="Binding site for ZF150-4"
                     /citation="[12]"
     misc_feature    143..151
                     /label=ZF6 BS
                     /note="Binding site for ZF150-4"
                     /citation="[13]"
     promoter        159..183
                     /label=YB_TATA Minimal Promoter
                     /citation="[6]"
     CDS             219..287
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="MDYKDHDGDYKDHDIDYKDDDDK"
     CDS             294..314
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             333..356
                     /codon_start=1
                     /label=Linker
                     /translation="GSGSGSGS"
     CDS             357..1031
                     /codon_start=1
                     /product="R95K non-fluorescent mutant of the noncytotoxic 
                     tetrameric variant of DsRed fluorescent protein"
                     /label=DsRed-Express2 R95K
                     /note="mammalian codon-optimized"
                     /citation="[1]"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWEKVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     CDS             1032..1103
                     /codon_start=1
                     /label=Linker
                     /translation="SGSGSGSGSGGGGSGGGGSGGGGS"
     CDS             1110..1223
                     /codon_start=1
                     /product="C-gp41-1 split intein fragment"
                     /label=intC
                     /citation="[4]"
                     /translation="MMLKKILKIEELDERELIDIEVSGNHLFYANDILTHNS"
     CDS             1230..1280
                     /codon_start=1
                     /label=Linker
                     /translation="GGGGSGGGGSGGGGSGT"
     CDS             1281..1544
                     /codon_start=1
                     /product="Zinc finger 43-8"
                     /label=ZF1
                     /label=ZF43-8
                     /citation="[14]"
                     /translation="ARPGERPFQCRICMRNFSRQDRLDRHTRTHTGEKPFQCRICMRNF
                     SQKEHLAGHLRTHTGEKPFQCRICMRNFSRRDNLNRHLKTHLR"
     CDS             1560..1580
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1593..1619
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1673..1897
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    1915..1939
                     /label=TUPV2 Barcode R
                     /note="TUPV barcode"
                     /citation="[7]"
     misc_feature    1963..2198
                     /label=cHS4
                     /note="Insulator sequence"
                     /citation="[2]"
     misc_feature    2302..2537
                     /label=cHS4
                     /note="Insulator sequence"
                     /citation="[3]"
     rep_origin      2666..3094
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      3164..3299
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3433..3554
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    3627..3643
                     /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(3651..3681)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3696..3717
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4005..4590)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4764..5621)
                     /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(5622..5726)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acgggcaaaa ccgatgttga cggactaatc ctgacaagat
       61 ctgcgaggtg taggggacgc gagtgtaggg gtgagctgtg tagggggact aagtgtaggg
      121 gacgcgagtg taggggtgag ctgtgtaggg gaacgctctc tagagggtat ataatggggg
      181 ccactagtct actaccagag ctcatcgcta gcgccaccat ggattataag gatcacgatg
      241 gcgactacaa ggaccacgat attgactaca aagacgatga cgataaaatg gctcctaaga
      301 aaaagcgcaa agtcggtatc catggcgttc ccggatcagg ctccggaagt ggctccatgg
      361 atagcactga gaacgtcatc aagcccttca tgcgcttcaa ggtgcacatg gagggctccg
      421 tgaacggcca cgagttcgag atcgagggcg agggcgaggg caagccctac gagggcaccc
      481 agaccgccaa gctccaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt
      541 ccccccagtt ccagtacggc tccaaggtgt acgtgaagca ccccgccgac atccccgact
      601 acaagaagct gtccttcccc gagggcttca agtgggagaa ggtgatgaac ttcgaggacg
      661 gcggcgtggt gaccgtgacc caggactcct ccctccagga cggcaccttc atctaccacg
      721 tgaagttcat cggcgtgaac ttcccctccg acggccccgt aatgcagaag aaaactctgg
      781 gctgggagcc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc
      841 acaaggcgct gaagctgaag ggcggcggcc actacctggt ggagttcaag tcaatctaca
      901 tggccaagaa gcccgtgaag ctgcccggct actactacgt ggactccaag ctggacatca
      961 cctcccacaa cgaggactac accgtggtgg agcagtacga gcgcgccgag gcccgccacc
     1021 acctgttcca gtctggttct ggttctggtt caggatccgg tggtggcggg tctggtggag
     1081 gaggtagtgg tggaggtggc tctgccacca tgatgctgaa gaagatcctg aagatcgagg
     1141 agctggacga gagagagctg atcgacatcg aggtgagcgg caaccacctg ttctacgcca
     1201 acgacatcct gacccacaac agcaacaggg gtggtggcgg gtctggtgga ggaggtagtg
     1261 gtggaggtgg ctctggtacc gctagacccg gggagcgccc cttccagtgt cgcatttgca
     1321 tgcggaactt ttcgcgccag gacaggcttg acaggcatac ccgtactcat accggtgaaa
     1381 aaccgtttca gtgtcggatc tgtatgcgaa atttctccca gaaggagcac ttggcggggc
     1441 atctacgtac gcacaccggc gagaagccat tccaatgccg aatatgcatg cgcaacttca
     1501 gtcgccgcga caacctgaac cggcacctaa aaacccacct gagagaattc ggctctggcc
     1561 ctaagaagaa gcgcaaagtt ggctctggca tgtacccata cgacgtacca gattacgctt
     1621 aggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     1681 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1741 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1801 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagtc
     1861 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcagg tcgaagaact
     1921 atttcctggc tgttacgcga gcgcatcgcg ataagctcta gagggacagc ccccccccaa
     1981 agcccccagg gatgtaatta cgtccctccc ccgctagggg gcagcagcga gccgcccggg
     2041 gctccgctcc ggtccggcgc tccccccgca tccccgagcc ggcagcgtgc ggggacagcc
     2101 cgggcacggg gaaggtggca cgggatcgct ttcctctgaa cgcttctcgc tgctctttga
     2161 gcctgcagac acctgggggg atacggggaa aaagctttag gctgaaagag agatttagaa
     2221 tgacagaatc atagaacggc ctgcgtacgg atggggctct agctctagag cttgatatcg
     2281 aattcctgca gccccgctag agggacagcc cccccccaaa gcccccaggg atgtaattac
     2341 gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg gtccggcgct
     2401 ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg aaggtggcac
     2461 gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca cctgggggga
     2521 tacggggaaa aagctttagg ctgaaagaga gatttagaat gacagaatca tagaacggcc
     2581 tgggcgcgcc tggtggcgaa cgatcaacag acgggagagg actaaagtct tcgctgacag
     2641 cgaccggact ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2701 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2761 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2821 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2881 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2941 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3001 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3061 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3121 gctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3181 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3241 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3301 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggaggtt gggcttcgga
     3361 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3421 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3481 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3541 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     3601 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3661 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3721 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3781 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3841 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3901 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3961 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4021 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4081 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4141 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4201 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4261 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4321 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4381 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4441 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4501 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag
     4561 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4621 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4681 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4741 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4801 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4861 gagggcttac catctggccc cagtgctgca atgataccgc gagatccacg ctcaccggct
     4921 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4981 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5041 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5101 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5161 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5221 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5281 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5341 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5401 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5461 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5521 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5581 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5641 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5701 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
