LOCUS       Exported                6985 bp ds-DNA     circular SYN 16-JUN-2019
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6985)
  AUTHORS   Viswajit_Kandula
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Feb 4, 2020 from SnapGene Viewer 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6985
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    31..55
                     /label=TUPV5 Barcode
     promoter        351..1532
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     intron          581..1523
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human
                     EF-1-alpha"
     regulatory      1633..1642
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
     CDS             1639..2355
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     misc_feature    1930..1938
                     /label=ZF13
                     /note="Binding site for ZF13-6 (Khalil, et al. Cell 2012)"
     misc_feature    1945..1953
                     /label=ZF14
                     /note="Binding site for ZF14-3 (Khalil, et al. Cell 2012)"
     CDS             2383..2439
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2452..2850
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    2901..3125
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3143..3167
                     /label=TUPV5 Barcode
     misc_feature    3191..3426
                     /label=cHS4
     misc_feature    3530..3765
                     /label=cHS4
     rep_origin      3894..4322
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      4392..4527
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4661..4782
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4831..4847)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4855..4871
                     /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(4879..4909)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_feature    4913..4921
                     /label=ZF55
                     /note="Binding site for ZF55-1 (Khalil, et al. Cell 2012)"
     protein_bind    4924..4945
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5233..5818)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5989..6849)
                     /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(6850..6954)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acggcagaaa ggtatggcac gcctaatctg gacacaagat
       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 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1681 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgag gggcgagggc
     1741 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1801 cccgtgccct ggcccaccct cgtgaccacc ctgagccacg gcgtgcagtg cttcgcccgc
     1861 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1921 caggagcgca ccatcttctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag
     1981 ttcgagggcg acaccctagt gaaccgcatc gagctgaagg gcgtcgactt caaggaggac
     2041 ggcaacatcc tggggcacaa gctggagtac aacttcaaca gccacaacat ctatatcatg
     2101 gccgtcaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa cgtggaggac
     2161 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2221 ctgctgcccg acagccacta cctgagcacc cagtccgtgc tgagcaaaga ccccaacgag
     2281 aagcgcgatc acatggtcct gctggagttc cgcaccgccg ccgggatcac tctcggcatg
     2341 gacgagctgt acaaggaatt cggtagcaag cttggtagtg gagcaactaa cttttccctg
     2401 ctcaaacaag ccggggatgt agaggagaac cccggccccg ccccccttaa gatggccaag
     2461 cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc
     2521 cccatctctg aggactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc
     2581 actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc
     2641 actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac
     2701 aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg
     2761 atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg
     2821 ctgccctctg gttatgtgtg ggagggctaa gcggccgctc gagtctagag ggcccgttta
     2881 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2941 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3001 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3061 ggacagcaag ggggaggatt gggaagtcaa tagcaggcat gctggggatg cggtgggctc
     3121 tatggcttct gaggcaggtc gatgcaccgt tcatatgtta tcgggacagc gcatcgcgat
     3181 aagctctaga gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc
     3241 gctagggggc agcagcgagc cgcccggggc tccgctccgg tccggcgctc cccccgcatc
     3301 cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg ggatcgcttt
     3361 cctctgaacg cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa
     3421 agctttaggc tgaaagagag atttagaatg acagaatcat agaacggcct gcgtacggat
     3481 ggggctctag ctctagagct tgatatcgaa ttcctgcagc cccgctagag ggacagcccc
     3541 cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc
     3601 gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg
     3661 gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc
     3721 tctttgagcc tgcagacacc tggggggata cggggaaaaa gctttaggct gaaagagaga
     3781 tttagaatga cagaatcata gaacggcctg ggcgcgcctg gtggcgaacg atcaacagac
     3841 gggagaggtg tgaagtcttc gctgacagcg accggactct agggggtatc cccacgcgcc
     3901 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3961 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4021 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4081 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4141 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4201 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4261 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4321 ttaattctgt ggaatgtgtg tcagttaggc tcaattagtc agcaaccata gtcccgcccc
     4381 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4441 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4501 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     4561 tatccatttt cggaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4621 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4681 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4741 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4801 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4861 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4921 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4981 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5041 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5101 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5161 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5221 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5281 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5341 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5401 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5461 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5521 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5581 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5641 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5701 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5761 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5821 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5881 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5941 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6001 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6061 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6121 gataccgcga gatccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6181 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6241 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6301 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6361 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6421 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6481 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6541 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6601 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6661 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6721 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6781 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6841 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6901 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6961 tccccgaaaa gtgccacctg acgtc
//
