LOCUS       Exported                5867 bp ds-DNA     circular SYN 01-MAR-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5867)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Sep 20, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5867
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    525..925
                     /label=Homology Arm 2
     protein_bind    549..560
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     misc_feature    950..2258
                     /label=TERT promoter
     regulatory      2284..2293
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2290..2847
                     /codon_start=1
                     /product="enhanced variant of secreted Gaussia luciferase"
                     /label=Gaussia-Dura Luc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     CDS             2290..2847
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     polyA_signal    2871..2992
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    3096..3495
                     /label=Homology Arm 1
     primer_bind     complement(3646..3662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3670..3686
                     /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(3694..3724)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3739..3760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4048..4636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4807..5667)
                     /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(5668..5772)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tgacgaccgc cggcgtttgc tgctagcttc cctgtccact agatggagag
      481 cttggctctg ttgggggaat tcatgaaagc accatctgtt taaacttcag ccttttgcag
      541 tttatcagga tgaggatgac cagcatgttg cccacaaaac caaagatgaa caccagtgag
      601 tagagcggag gcaggaggcg ggctgcgatt tgcttcacat tgattttttg gcagggctcc
      661 gatgtataat aattgatgtc atagattgga cttgacactt gataatccat cttgttccac
      721 cctgtgcata aataaaaagt gatcttttat aaagtcctag aatgtattta gttgccctcc
      781 atgaatgcaa actgttttat acatcaatag gtttttaatt gcctacatag atgtctacat
      841 tgaattaact ctctttttgg ccaagcaatg aagttttgta gtgaagggaa ggtttgctgc
      901 tagcttccct gtccactaga tggagaagct tgttaacacg cgtagatctc agcccaggga
      961 gggtgcgagg cctgttcaaa tgctagctcc ataaataaag caatttcctc cggcagtttc
     1021 tgaaagtagg aaaggttaca tttaaggttg cgtttgttag catttcagtg tttgccgacc
     1081 tcagctacag catccctgca aggcctcggg agacccagaa gtttctcgcc ccttagatcc
     1141 aaacttgagc aacccggagt ctggattcct gggaagtcct cagctgtcct gcggttgtgc
     1201 cggggcccca ggtctggagg ggaccagtgg ccgtgtggct tctactgctg ggctggaagt
     1261 cgggcctcct agctctgcag tccgaggctt ggagccaggt gcctggaccc cgaggctgcc
     1321 ctccaccctg tgcgggcggg atgtgaccag atgttggcct catctgccag acagagtgcc
     1381 ggggcccagg gtcaaggccg ttgtggctgg tgtgaggcgc ccggtgcgcg gccagcagga
     1441 gcgcctggct ccatttccca ccctttctcg acgggaccgc cccggtgggt gattaacaga
     1501 tttggggtgg tttgctcatg gtggggaccc ctcgccgcct gagaacctgc aaagagaaat
     1561 gacgggcctg tgtcaaggag cccaagtcgc ggggaagtgt tgcagggagg cactccggga
     1621 ggtcccgcgt gcccgtccag ggagcaatgc gtcctcgggt tcgtccccag ccgcgtctac
     1681 gcgcctccgt cctccccttc acgtccggca ttcgtggtgc ccggagcccg acgccccgcg
     1741 tccggacctg gaggcagccc tgggtctccg gatcaggcca gcggccaaag ggtcgccgca
     1801 cgcacctgtt cccagggcct ccacatcatg gcccctccct cgggttaccc cacagcctag
     1861 gccgattcga cctctctccg ctggggccct cgctggcgtc cctgcaccct gggagcgcga
     1921 gcggcgcgcg ggcggggaag cgcggcccag acccccgggt ccgcccggag cagctgcgct
     1981 gtcggggcca ggccgggctc ccagtggatt cgcgggcaca gacgcccagg accgcgcttc
     2041 ccacgtggcg gagggactgg ggacccgggc acccgtcctg ccccttcacc ttccagctcc
     2101 gcctcctccg cgcggacccc gccccgtccc gacccctccc gggtccccgg cccagccccc
     2161 tccgggccct cccagcccct ccccttcctt tccgcggccc cgccctctcc tcgcggcgcg
     2221 agtttcaggc agcgctgcgt cctgctgcgc acgtgggaaa gcttggtacc gagctcggat
     2281 ccagccacca tgggagtcaa agttctgttt gccctgatct gcatcgctgt ggccgaggcc
     2341 aagcccaccg agaacaacga agacttcaac atcgtggccg tggccagcaa cttcgcgacc
     2401 acggatctcg atgctgaccg cgggaagttg cccggcaaga agctgccgct ggaggtgctc
     2461 aaagagctgg aagccaatgc ccggaaagct ggctgcacca ggggctgtct gatctgcctg
     2521 tcccacatca agtgcacgcc caagatgaag aagttcatcc caggacgctg ccacacctac
     2581 gaaggcgaca aagagtccgc acagggcggc ataggcgagg cgatcgtcga cattcctgag
     2641 attcctgggt tcaaggactt ggagcccctg gagcagttca tcgcacaggt cgatctgtgt
     2701 gtggactgca caactggctg cctcaaaggg cttgccaacg tgcagtgttc tgacctgctc
     2761 aagaagtggc tgccgcaacg ctgtgcgacc tttgccagca agatccaggg ccaggtggac
     2821 aagatcaagg gggccggtgg tgactaagcg gccgcttcga gcagacatga taagatacat
     2881 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2941 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3001 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     3061 gtaaaacctc tacaaatgtg gtactcgaga cgcgtgctct gttgggggaa ttcatgaaag
     3121 caccatctca ccaaataaaa tcttgtgctc tatagcacca tggagtgaat gaagctttga
     3181 caacaatttt aagaaactgg gtcaagcatg gtgcagctca gcctttttgt tgttgttgtt
     3241 gttgttgtga gacagggtct cactctgtcg cccaggctgg agtgcagtgg cacgatcatg
     3301 gctcactgca gcctggacca tcccgggctc aagcaatcct ctcacctcac ccctccaagt
     3361 ggctggggac tacaggcgcc accacaccaa gctaattttg tttattttta tagatacaag
     3421 atcacactat gttgcccagg ctggtctcga actcctgggc tcaagcgatc ctcctgcctc
     3481 agcctcctag aatgcattta aattttgctg ctagcttccc tgtccactag atggagagct
     3541 tggctctgtt gggggaattc atgaaagcac catctgcggc cgcgtcgtat cggatcccgg
     3601 gcccgtcgac tgcagaggcc tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     3661 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3721 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3781 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3841 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3901 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3961 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4021 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4081 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4141 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4201 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4261 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4321 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4381 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4441 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4501 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4561 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4621 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4681 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4741 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4801 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4861 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4921 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4981 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5041 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5101 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5161 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5221 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5281 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5341 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5401 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5461 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5521 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5581 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5641 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5701 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5761 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     5821 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
