LOCUS       Exported                6370 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 6370)
  TITLE     Hermann, et al. 2014
REFERENCE   2  (bases 1 to 6370)
  TITLE     Bintu et al., 2016
REFERENCE   3  (bases 1 to 6370)
  TITLE     Bintu et al., 2016
REFERENCE   4  (bases 1 to 6370)
  TITLE     Beerli et al., 1998
REFERENCE   5  (bases 1 to 6370)
  TITLE     Xu et al., 2009, Donahue et al., 2020
REFERENCE   6  (bases 1 to 6370)
  TITLE     Xu et al., 2009, Donahue et al., 2020
REFERENCE   7  (bases 1 to 6370)
  AUTHORS   Leonard Lab
  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COMMENT     SGRef: number: 1; type: ""Unpublished Work""
            SGRef: number: 2; type: ""Unpublished Work""
            SGRef: number: 3; type: ""Unpublished Work""
            SGRef: number: 4; type: ""Unpublished Work""
            SGRef: number: 5; type: ""Unpublished Work""
            SGRef: number: 6; type: ""Unpublished Work""
FEATURES             Location/Qualifiers
     source          1..6370
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    31..55
                     /label=TUPV2 Barcode F
                     /note="TUPV barcode"
                     /citation="[5]"
     promoter        351..1532
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human
                     elongationfactor EF-1-alpha"
     CDS             1639..1707
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="MDYKDHDGDYKDHDIDYKDDDDK"
     CDS             1714..1734
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1750..1899
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation
                     domain of herpes simplex virus VP16"
                     /label=VP64 AD
                     /label=VP64
                     /citation="[4]"
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             1900..1953
                     /codon_start=1
                     /label=Linker
                     /translation="GSGGGGSGGGGTGGGGTG"
     CDS             1954..1968
                     /codon_start=1
                     /product="Five amino acid motif upstream of intN"
                     /label=TRSGY
                     /note="At the C-terminus of the adjoining extein"
                     /translation="TRSGY"
     CDS             1969..2232
                     /codon_start=1
                     /product="N-gp41-1 split intein fragment"
                     /label=intN
                     /citation="[1]"
                     /translation="CLDLKTQVQTPQGMKEISNIQVGDLVLSNTGYNEVLNVFPKSKKK
                     SYKITLEDGKEIICSEEHLFPTQTGEMNISGGLKEGMCLYVKE"
     polyA_signal    2286..2510
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    2528..2552
                     /label=TUPV2 Barcode R
                     /note="TUPV barcode"
                     /citation="[6]"
     misc_feature    2576..2811
                     /label=cHS4
                     /note="Insulator sequence"
                     /citation="[2]"
     misc_feature    2915..3150
                     /label=cHS4
                     /note="Insulator sequence"
                     /citation="[3]"
     rep_origin      3279..3707
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      3777..3912
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4046..4167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    4240..4256
                     /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(4264..4294)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4309..4330
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4618..5203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5377..6234)
                     /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(6235..6339)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acgggcaaaa ccgatgttga cggactaatc ctgacaagat
       61 ctggggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
      121 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      181 gggccagata tacgcgtgag taattcatac aaaaggactc gcccctgcct tggggaatcc
      241 cagggaccgt cgttaaactc ccactaacgt agaacccaga gatcgctgcg ttcccgcccc
      301 ctcacccgcc cgctctcgtc atcactgagg tggagaagag catgcgtgag gctccggtgc
      361 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
      421 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
      481 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
      541 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
      601 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacgcc
      661 cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt
      721 cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcttgggc
      781 gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata
      841 agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata
      901 gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg
      961 gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc
     1021 caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg
     1081 cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt
     1141 gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc
     1201 gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag
     1261 ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct
     1321 cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc
     1381 cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct
     1441 tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc
     1501 aaagtttttt tcttccattt caggtgtcgt gagagctcgt ttagtgaacc gtcagatcgc
     1561 ctggagacgc catccacgct gttttgacct ccatagaaga gaacgggacc gatccaaatt
     1621 ctcgaggcta gcgccaccat ggactataaa gaccatgacg gtgactacaa ggatcacgac
     1681 atagattata aagacgatga cgacaagatg gcacctaaga agaagcgcaa agttggctct
     1741 ggcgaattcg acgcccttga tgactttgac ttggacatgc tggggtcaga cgcactcgat
     1801 gattttgacc ttgatatgct gggaagcgat gcacttgatg actttgactt ggacatgctc
     1861 ggctccgacg ctcttgatga tttcgatctt gacatgctcg gatccggtgg tggcgggtct
     1921 ggaggaggag gtactggtgg aggtggcact ggtaccagaa gcggatactg cctggacctg
     1981 aagacccagg tgcagacccc tcagggcatg aaggagatca gcaacatcca ggtgggcgac
     2041 ctggtgctga gcaacaccgg ctacaacgag gtgctgaacg tgttccccaa gagcaagaag
     2101 aagagctaca agatcaccct ggaggacggc aaggagatca tctgcagcga ggagcacctg
     2161 ttccccaccc agaccggcga gatgaacatc agcggcggcc tgaaggaggg catgtgcctg
     2221 tacgtgaagg agtgagcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc
     2281 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2341 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2401 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2461 ggattgggaa gtcaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2521 aggtcgaaga actatttcct ggctgttacg cgagcgcatc gcgataagct ctagagggac
     2581 agcccccccc caaagccccc agggatgtaa ttacgtccct cccccgctag ggggcagcag
     2641 cgagccgccc ggggctccgc tccggtccgg cgctcccccc gcatccccga gccggcagcg
     2701 tgcggggaca gcccgggcac ggggaaggtg gcacgggatc gctttcctct gaacgcttct
     2761 cgctgctctt tgagcctgca gacacctggg gggatacggg gaaaaagctt taggctgaaa
     2821 gagagattta gaatgacaga atcatagaac ggcctgcgta cggatggggc tctagctcta
     2881 gagcttgata tcgaattcct gcagccccgc tagagggaca gccccccccc aaagccccca
     2941 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     3001 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     3061 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     3121 acacctgggg ggatacgggg aaaaagcttt aggctgaaag agagatttag aatgacagaa
     3181 tcatagaacg gcctgggcgc gcctggtggc gaacgatcaa cagacgggag aggactaaag
     3241 tcttcgctga cagcgaccgg actctagggg gtatccccac gcgccctgta gcggcgcatt
     3301 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3361 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3421 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3481 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3541 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3601 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3661 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3721 gtgtgtcagt taggctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3781 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3841 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     3901 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggag
     3961 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4021 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4081 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4141 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4201 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4261 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4321 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4381 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4441 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4501 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4561 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4621 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4681 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4741 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4801 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4861 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4921 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4981 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5041 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5101 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     5161 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5221 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5281 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5341 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5401 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5461 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagatcc
     5521 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5581 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5641 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5701 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5761 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5821 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5881 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5941 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6001 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6061 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6121 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6181 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6241 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6301 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6361 acctgacgtc
//
