LOCUS       Exported                6340 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 6340)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   2  (bases 1 to 6340)
  TITLE     Bintu et al., 2016
REFERENCE   3  (bases 1 to 6340)
  TITLE     Bintu et al., 2016
REFERENCE   4  (bases 1 to 6340)
  TITLE     Xu et al., 2009, Donahue et al., 2020
REFERENCE   5  (bases 1 to 6340)
  TITLE     Xu et al., 2009, Donahue et al., 2020
REFERENCE   6  (bases 1 to 6340)
  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COMMENT     SGRef: number: 1; type: ""Unpublished Work""
            SGRef: number: 2; type: ""Unpublished Work""
            SGRef: number: 3; type: ""Unpublished Work""
            SGRef: number: 4; type: ""Unpublished Work""
            SGRef: number: 5; type: ""Unpublished Work""
FEATURES             Location/Qualifiers
     source          1..6340
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    31..55
                     /label=TUPV2 Barcode F
                     /note="TUPV barcode"
                     /citation="[4]"
     enhancer        199..578
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        579..782
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        827..845
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             871..939
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="MDYKDHDGDYKDHDIDYKDDDDK"
     CDS             946..966
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             973..1242
                     /codon_start=1
                     /product="Zinc finger 37-12"
                     /label=ZF2
                     /label=ZF37-12
                     /citation="[1]"
                     /translation="ARPGERPFQCRICMRNFSRNFILQRHTRTHTGEKPFQCRICMRNF
                     SDRANLRRHLRTHTGEKPFQCRICMRNFSRHDQLTRHLKTHLRGS"
     CDS             1243..1299
                     /codon_start=1
                     /label=Linker
                     /translation="GSGGGGSGGGGSGGGGSGT"
     CDS             1300..2202
                     /codon_start=1
                     /product="Truncated ABA insensitive 1 protein"
                     /label=ABI1
                     /translation="MTRVPLYGFTSICGRRPEMEAAVSTIPRFLQSSSGSMLDGRFDPQ
                     SAAHFFGVYDGHGGSQVANYCRERMHLALAEEIAKEKPMLCDGDTWLEKWKKALFNSFL
                     RVDSEIESVAPETVGSTSVVAVVFPSHIFVANCGDSRAVLCRGKTALPLSVDHKPDRED
                     EAARIEAAGGKVIQWNGARVFGVLAMSRSIGDRYLKPSIIPDPEVTAVKRVKEDDCLIL
                     ASDGVWDVMTDEEACEMARKRILLWHKKNAVAGDASLLADERRKEGKDPAAMSAAEYLS
                     KLAIQRGSKDNISVVVVDLK"
     polyA_signal    2256..2480
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    2498..2522
                     /label=TUPV2 Barcode R
                     /note="TUPV barcode"
                     /citation="[5]"
     misc_feature    2546..2781
                     /label=cHS4
                     /note="Insulator sequence"
                     /citation="[2]"
     misc_feature    2885..3120
                     /label=cHS4
                     /note="Insulator sequence"
                     /citation="[3]"
     rep_origin      3249..3677
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      3747..3882
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4016..4137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    4210..4226
                     /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(4234..4264)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4279..4300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4588..5173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5347..6204)
                     /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(6205..6309)
                     /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 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      241 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      301 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      361 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      421 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      481 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      541 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      601 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      661 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      721 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      781 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
      841 atagggagtc ccaagctggc tagcgccacc atggactaca aagaccacga cggcgattac
      901 aaggatcatg acatcgatta caaagacgat gacgataaga tggctcctaa gaaaaagcgc
      961 aaagtctccg gagctagacc cggggagcgc cccttccagt gtcgcatttg catgcggaac
     1021 ttttcgagaa atttcatttt gcaaagacat acccgtactc ataccggtga aaaaccgttt
     1081 cagtgtcgga tctgtatgcg aaatttctcc gatagagcta atttgaggag gcatctgcgt
     1141 acgcacaccg gcgagaagcc attccaatgc cgaatatgca tgcgcaactt cagtagacat
     1201 gatcaattga ctagacacct aaaaacccac ctgagaggat ctggatccgg tggtggcggg
     1261 tctggaggag gaggtagtgg tggaggtggc tctggtacca tgacgcgggt gcctttgtat
     1321 ggttttactt cgatttgtgg aagaaggcct gagatggaag ctgctgtttc gactatacca
     1381 agattccttc aatcttcctc tggttcgatg ttagatggtc ggtttgatcc tcaatccgcc
     1441 gctcatttct tcggtgttta cgacggccat ggcggttctc aggtagcgaa ctattgtaga
     1501 gagaggatgc atttggcttt ggcggaggag atagctaagg agaaaccgat gctctgcgat
     1561 ggtgatacgt ggctggagaa gtggaagaaa gctcttttca actcgttcct gagagttgac
     1621 tcggagattg agtcagttgc gccggagacg gttgggtcaa cgtcggtggt tgccgttgtt
     1681 ttcccgtctc acatcttcgt cgctaactgc ggtgactcta gagccgttct ttgccgcggc
     1741 aaaactgcac ttccattatc cgttgaccat aaaccggata gagaagatga agctgcgagg
     1801 attgaagccg caggagggaa agtgattcag tggaatggag ctcgtgtttt cggtgttctc
     1861 gccatgtcga gatccattgg cgatagatac ttgaaaccat ccatcattcc tgatcctgaa
     1921 gtgacggctg tgaagagagt aaaagaagat gattgtctga ttttggcgag tgacggggtt
     1981 tgggatgtaa tgacggatga agaagcgtgt gagatggcaa ggaagcggat tctcttgtgg
     2041 cacaagaaaa acgcggtggc tggggatgca tcgttgctcg cggatgagcg gagaaaggaa
     2101 gggaaagatc ctgcggcgat gtccgcggct gagtatttgt caaagctggc gatacagaga
     2161 ggaagcaaag acaacataag tgtggtggtg gttgatttga agtaggcggc cgctcgagtc
     2221 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2281 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2341 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2401 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gtcaatagca ggcatgctgg
     2461 ggatgcggtg ggctctatgg cttctgaggc aggtcgaaga actatttcct ggctgttacg
     2521 cgagcgcatc gcgataagct ctagagggac agcccccccc caaagccccc agggatgtaa
     2581 ttacgtccct cccccgctag ggggcagcag cgagccgccc ggggctccgc tccggtccgg
     2641 cgctcccccc gcatccccga gccggcagcg tgcggggaca gcccgggcac ggggaaggtg
     2701 gcacgggatc gctttcctct gaacgcttct cgctgctctt tgagcctgca gacacctggg
     2761 gggatacggg gaaaaagctt taggctgaaa gagagattta gaatgacaga atcatagaac
     2821 ggcctgcgta cggatggggc tctagctcta gagcttgata tcgaattcct gcagccccgc
     2881 tagagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc ccccgctagg
     2941 gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg catccccgag
     3001 ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg ctttcctctg
     3061 aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg aaaaagcttt
     3121 aggctgaaag agagatttag aatgacagaa tcatagaacg gcctgggcgc gcctggtggc
     3181 gaacgatcaa cagacgggag aggactaaag tcttcgctga cagcgaccgg actctagggg
     3241 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3301 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3361 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3421 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3481 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3541 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3601 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3661 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt taggctcaat tagtcagcaa
     3721 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3781 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3841 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3901 tcccgggagc ttgtatatcc attttcggag gttgggcttc ggaatcgttt tccgggacgc
     3961 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4021 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4081 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4141 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     4201 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4261 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4321 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4381 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4441 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4501 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4561 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4621 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4681 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4741 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4801 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4861 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4921 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4981 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5041 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5101 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5161 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5221 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5281 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5341 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5401 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5461 ccccagtgct gcaatgatac cgcgagatcc acgctcaccg gctccagatt tatcagcaat
     5521 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5581 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5641 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5701 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5761 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5821 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5881 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5941 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6001 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6061 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6121 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6181 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6241 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6301 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
