LOCUS       Exported                6421 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 6421)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   2  (bases 1 to 6421)
  TITLE     Hermann, et al. 2014
REFERENCE   3  (bases 1 to 6421)
  TITLE     Bintu et al., 2016
REFERENCE   4  (bases 1 to 6421)
  TITLE     Bintu et al., 2016
REFERENCE   5  (bases 1 to 6421)
  TITLE     Xu et al., 2009, Donahue et al., 2020
REFERENCE   6  (bases 1 to 6421)
  TITLE     Xu et al., 2009, Donahue et al., 2020
REFERENCE   7  (bases 1 to 6421)
  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COMMENT     SGRef: number: 1; type: ""Unpublished Work""
            SGRef: number: 2; type: ""Unpublished Work""
            SGRef: number: 3; type: ""Unpublished Work""
            SGRef: number: 4; type: ""Unpublished Work""
            SGRef: number: 5; type: ""Unpublished Work""
            SGRef: number: 6; type: ""Unpublished Work""
FEATURES             Location/Qualifiers
     source          1..6421
                     /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..1656
                     /codon_start=1
                     /product="Five amino acid motif upstream of intN"
                     /label=TRSGY
                     /note="At the C-terminus of the adjoining extein"
                     /translation="MTRSGY"
     CDS             1657..1920
                     /codon_start=1
                     /product="N-gp41-1 split intein fragment"
                     /label=intN
                     /citation="[2]"
                     /translation="CLDLKTQVQTPQGMKEISNIQVGDLVLSNTGYNEVLNVFPKSKKK
                     SYKITLEDGKEIICSEEHLFPTQTGEMNISGGLKEGMCLYVKE"
     CDS             1921..1959
                     /codon_start=1
                     /label=Linker
                     /translation="TGSGSGSGSGTGG"
     CDS             1960..2229
                     /codon_start=1
                     /product="Zinc finger 13-6"
                     /label=ZF10
                     /label=ZF13-6
                     /citation="[1]"
                     /translation="ARPGERPFQCRICMRNFSTNQKLEVHTRTHTGEKPFQCRICMRNF
                     SVRHNLQRHLRTHTGEKPFQCRICMRNFSQHPNLTRHLKTHLRGS"
     CDS             2230..2250
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2257..2283
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2337..2561
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    2579..2603
                     /label=TUPV2 Barcode R
                     /note="TUPV barcode"
                     /citation="[6]"
     misc_feature    2627..2862
                     /label=cHS4
                     /note="Insulator sequence"
                     /citation="[3]"
     misc_feature    2966..3201
                     /label=cHS4
                     /note="Insulator sequence"
                     /citation="[4]"
     rep_origin      3330..3758
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      3828..3963
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4097..4218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    4291..4307
                     /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(4315..4345)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4360..4381
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4669..5254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5428..6285)
                     /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(6286..6390)
                     /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 gaccagaagc ggatactgcc tggacctgaa gacccaggtg
     1681 cagacccctc agggcatgaa ggagatcagc aacatccagg tgggcgacct ggtgctgagc
     1741 aacaccggct acaacgaggt gctgaacgtg ttccccaaga gcaagaagaa gagctacaag
     1801 atcaccctgg aggacggcaa ggagatcatc tgcagcgagg agcacctgtt ccccacccag
     1861 accggcgaga tgaacatcag cggcggcctg aaggagggca tgtgcctgta cgtgaaggag
     1921 actggttctg gttctggttc aggatccggt accggtggag ctagacccgg ggagcgcccc
     1981 ttccagtgtc gcatttgcat gcggaacttt tcgactaatc aaaaattgga agttcatacc
     2041 cgtactcata ccggtgaaaa accgtttcag tgtcggatct gtatgcgaaa tttctccgtt
     2101 agacataatt tgcaaagaca tctacgtacg cacaccggcg agaagccatt ccaatgccga
     2161 atatgcatgc gcaacttcag tcaacatcca aatttgacta gacacctaaa aacccacctg
     2221 agaggatctc ctaagaagaa gcgcaaagtt ggcatgtacc catacgacgt accagattac
     2281 gcttaggcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     2341 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2401 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2461 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2521 agtcaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg caggtcgaag
     2581 aactatttcc tggctgttac gcgagcgcat cgcgataagc tctagaggga cagccccccc
     2641 ccaaagcccc cagggatgta attacgtccc tcccccgcta gggggcagca gcgagccgcc
     2701 cggggctccg ctccggtccg gcgctccccc cgcatccccg agccggcagc gtgcggggac
     2761 agcccgggca cggggaaggt ggcacgggat cgctttcctc tgaacgcttc tcgctgctct
     2821 ttgagcctgc agacacctgg ggggatacgg ggaaaaagct ttaggctgaa agagagattt
     2881 agaatgacag aatcatagaa cggcctgcgt acggatgggg ctctagctct agagcttgat
     2941 atcgaattcc tgcagccccg ctagagggac agcccccccc caaagccccc agggatgtaa
     3001 ttacgtccct cccccgctag ggggcagcag cgagccgccc ggggctccgc tccggtccgg
     3061 cgctcccccc gcatccccga gccggcagcg tgcggggaca gcccgggcac ggggaaggtg
     3121 gcacgggatc gctttcctct gaacgcttct cgctgctctt tgagcctgca gacacctggg
     3181 gggatacggg gaaaaagctt taggctgaaa gagagattta gaatgacaga atcatagaac
     3241 ggcctgggcg cgcctggtgg cgaacgatca acagacggga gaggactaaa gtcttcgctg
     3301 acagcgaccg gactctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3361 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3421 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3481 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3541 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3601 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3661 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3721 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3781 ttaggctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3841 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3901 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3961 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga ggttgggctt
     4021 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4081 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4141 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4201 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     4261 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4321 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4381 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4441 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4501 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4561 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4621 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4681 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4741 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4801 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4861 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4921 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4981 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5041 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5101 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5161 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa
     5221 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5281 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5341 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5401 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5461 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5521 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagatc cacgctcacc
     5581 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5641 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5701 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5761 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5821 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5881 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5941 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6001 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6061 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6121 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6181 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6241 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6301 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6361 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6421 c
//
