LOCUS       Exported                5184 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 5184)
  TITLE     Bintu et al., 2016
REFERENCE   2  (bases 1 to 5184)
  TITLE     Bintu et al., 2016
REFERENCE   3  (bases 1 to 5184)
  TITLE     Beerli et al., 1998)
REFERENCE   4  (bases 1 to 5184)
  TITLE     Hermann, et al. 2014
REFERENCE   5  (bases 1 to 5184)
  TITLE     Xu et al., 2009, Donahue et al., 2020
REFERENCE   6  (bases 1 to 5184)
  TITLE     Hansen et al., 2014
REFERENCE   7  (bases 1 to 5184)
  TITLE     Xu et al., 2009, Donahue et al., 2020
REFERENCE   8  (bases 1 to 5184)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   9  (bases 1 to 5184)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   10  (bases 1 to 5184)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   11  (bases 1 to 5184)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   12  (bases 1 to 5184)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   13  (bases 1 to 5184)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   14  (bases 1 to 5184)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   15  (bases 1 to 5184)
  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COMMENT     SGRef: number: 1; type: ""Unpublished Work""
            SGRef: number: 2; type: ""Unpublished Work""
            SGRef: number: 3; type: ""Unpublished Work""
            SGRef: number: 4; type: ""Unpublished Work""
            SGRef: number: 5; type: ""Unpublished Work""
            SGRef: number: 6; type: ""Unpublished Work""
            SGRef: number: 7; type: ""Unpublished Work""
            SGRef: number: 8; type: ""Unpublished Work""
            SGRef: number: 9; type: ""Unpublished Work""
            SGRef: number: 10; type: ""Unpublished Work""
            SGRef: number: 11; type: ""Unpublished Work""
            SGRef: number: 12; type: ""Unpublished Work""
            SGRef: number: 13; type: ""Unpublished Work""
            SGRef: number: 14; type: ""Unpublished Work""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..5184
                     /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=ZF2 BS
                     /note="Binding site for ZF37-12"
                     /citation="[8]"
     misc_feature    83..91
                     /label=ZF2 BS
                     /note="Binding site for ZF37-12"
                     /citation="[9]"
     misc_feature    98..106
                     /label=ZF2 BS
                     /note="Binding site for ZF37-12"
                     /citation="[10]"
     misc_feature    113..121
                     /label=ZF2 BS
                     /note="Binding site for ZF37-12"
                     /citation="[11]"
     misc_feature    128..136
                     /label=ZF2 BS
                     /note="Binding site for ZF37-12"
                     /citation="[12]"
     misc_feature    143..151
                     /label=ZF2 BS
                     /note="Binding site for ZF37-12"
                     /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             330..479
                     /codon_start=1
                     /locus_tag="VP64"
                     /product="tetrameric repeat of the minimal activation
                     domain of herpes simplex virus VP16"
                     /label=VP64 AD
                     /citation="[3]"
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             480..530
                     /codon_start=1
                     /label=Linker
                     /translation="GSGGGGSGGGGSGGGGS"
     CDS             537..650
                     /codon_start=1
                     /product="C-gp41-1 split intein fragment"
                     /label=intC
                     /citation="[4]"
                     /translation="MMLKKILKIEELDERELIDIEVSGNHLFYANDILTHNS"
     CDS             657..707
                     /codon_start=1
                     /label=Linker
                     /translation="GGGGSGGGGSGGGGSGT"
     CDS             708..971
                     /codon_start=1
                     /product="Zinc finger 43-8"
                     /label=ZF1
                     /label=ZF43-8
                     /citation="[14]"
                     /translation="ARPGERPFQCRICMRNFSRQDRLDRHTRTHTGEKPFQCRICMRNF
                     SQKEHLAGHLRTHTGEKPFQCRICMRNFSRRDNLNRHLKTHLR"
     CDS             987..1007
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1020..1046
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1100..1324
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    1342..1366
                     /label=TUPV2 Barcode R
                     /note="TUPV barcode"
                     /citation="[7]"
     misc_feature    1390..1625
                     /label=cHS4
                     /note="Insulator sequence"
                     /citation="[1]"
     misc_feature    1729..1964
                     /label=cHS4
                     /note="Insulator sequence"
                     /citation="[2]"
     rep_origin      2093..2521
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      2591..2726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2860..2981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    3054..3070
                     /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(3078..3108)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3123..3144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3432..4017)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4191..5048)
                     /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(5049..5153)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acgggcaaaa ccgatgttga cggactaatc ctgacaagat
       61 ctgcgaggag gacgtgacgc gagaggacgt gtgagctgag gacgtggact aagaggacgt
      121 gacgcgagag gacgtgtgag ctgaggacgt gaacgctctc tagagggtat ataatggggg
      181 ccactagtct actaccagag ctcatcgcta gcgccaccat ggactataaa gaccatgacg
      241 gtgactacaa ggatcacgac atagattata aagacgatga cgacaagatg gcacctaaga
      301 agaagcgcaa agttggctct ggcgaattcg acgcccttga tgactttgac ttggacatgc
      361 tggggtcaga cgcactcgat gattttgacc ttgatatgct gggaagcgat gcacttgatg
      421 actttgactt ggacatgctc ggctccgacg ctcttgatga tttcgatctt gacatgctcg
      481 gatccggtgg tggcgggtct ggtggaggag gtagtggtgg aggtggctct gccaccatga
      541 tgctgaagaa gatcctgaag atcgaggagc tggacgagag agagctgatc gacatcgagg
      601 tgagcggcaa ccacctgttc tacgccaacg acatcctgac ccacaacagc aacaggggtg
      661 gtggcgggtc tggtggagga ggtagtggtg gaggtggctc tggtaccgct agacccgggg
      721 agcgcccctt ccagtgtcgc atttgcatgc ggaacttttc gcgccaggac aggcttgaca
      781 ggcatacccg tactcatacc ggtgaaaaac cgtttcagtg tcggatctgt atgcgaaatt
      841 tctcccagaa ggagcacttg gcggggcatc tacgtacgca caccggcgag aagccattcc
      901 aatgccgaat atgcatgcgc aacttcagtc gccgcgacaa cctgaaccgg cacctaaaaa
      961 cccacctgag agaattcggc tctggcccta agaagaagcg caaagttggc tctggcatgt
     1021 acccatacga cgtaccagat tacgcttagg cggccgctcg agtctagagg gcccgtttaa
     1081 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     1141 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     1201 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     1261 gacagcaagg gggaggattg ggaagtcaat agcaggcatg ctggggatgc ggtgggctct
     1321 atggcttctg aggcaggtcg aagaactatt tcctggctgt tacgcgagcg catcgcgata
     1381 agctctagag ggacagcccc cccccaaagc ccccagggat gtaattacgt ccctcccccg
     1441 ctagggggca gcagcgagcc gcccggggct ccgctccggt ccggcgctcc ccccgcatcc
     1501 ccgagccggc agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg gatcgctttc
     1561 ctctgaacgc ttctcgctgc tctttgagcc tgcagacacc tggggggata cggggaaaaa
     1621 gctttaggct gaaagagaga tttagaatga cagaatcata gaacggcctg cgtacggatg
     1681 gggctctagc tctagagctt gatatcgaat tcctgcagcc ccgctagagg gacagccccc
     1741 ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag cagcgagccg
     1801 cccggggctc cgctccggtc cggcgctccc cccgcatccc cgagccggca gcgtgcgggg
     1861 acagcccggg cacggggaag gtggcacggg atcgctttcc tctgaacgct tctcgctgct
     1921 ctttgagcct gcagacacct ggggggatac ggggaaaaag ctttaggctg aaagagagat
     1981 ttagaatgac agaatcatag aacggcctgg gcgcgcctgg tggcgaacga tcaacagacg
     2041 ggagaggact aaagtcttcg ctgacagcga ccggactcta gggggtatcc ccacgcgccc
     2101 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2161 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2221 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2281 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2341 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2401 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2461 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2521 taattctgtg gaatgtgtgt cagttaggct caattagtca gcaaccatag tcccgcccct
     2581 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2641 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     2701 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     2761 atccattttc ggaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     2821 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     2881 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2941 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     3001 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     3061 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3121 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3181 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3241 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3301 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3361 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3421 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3481 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3541 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3601 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3661 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3721 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3781 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3841 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3901 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3961 ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4021 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4081 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4141 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4201 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4261 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4321 ataccgcgag atccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4381 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4441 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4501 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4561 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4621 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4681 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4741 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4801 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4861 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4921 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4981 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5041 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5101 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5161 ccccgaaaag tgccacctga cgtc
//
