LOCUS       Exported                7416 bp ds-DNA     circular SYN 25-MAR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7416)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, May 10, 2021 from SnapGene Viewer 5.2.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7416
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      33..42
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             39..695
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MASPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             723..743
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             756..1472
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    3125..3180
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3539..3555)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3563..3579
                     /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(3587..3617)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3632..3653
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3953..4541)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4712..5572)
                     /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(5573..5677)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5707..6086
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6089..6364
                     /label=chicken beta-actin promoter
     intron          6365..7373
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
ORIGIN
        1 gcaaagaatt gcggccgtct caggccgaat tcgccaccat ggcatcccct atactaggtt
       61 attggaaaat taagggcctt gtgcaaccca ctcgacttct tttggaatat cttgaagaaa
      121 aatatgaaga gcatttgtat gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg
      181 aattgggttt ggagtttccc aatcttcctt attatattga tggtgatgtt aaattaacac
      241 agtctatggc catcatacgt tatatagctg acaagcacaa catgttgggt ggttgtccaa
      301 aagagcgtgc agagatttca atgcttgaag gagcggtttt ggatattaga tacggtgttt
      361 cgagaattgc atatagtaaa gactttgaaa ctctcaaagt tgattttctt agcaagctac
      421 ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa aacatattta aatggtgatc
      481 atgtaaccca tcctgacttc atgttgtatg acgctcttga tgttgtttta tacatggacc
      541 caatgtgcct ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt gaagctatcc
      601 cacaaattga taagtacttg aaatccagca agtatatagc atggcctttg cagggctggc
      661 aagccacgtt tggtggtggc gaccatcctc caaaatcgga tgattacgat atcccaacga
      721 ccgaaaacct gtattttcag ggtgtcgacg gtaccatggt gagcaagggc gaggagctgt
      781 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
      841 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
      901 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
      961 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     1021 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1081 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1141 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     1201 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     1261 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1321 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga
     1381 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1441 ggatcactct cggcatggac gagctgtaca agggtaccat ggcttcccca tctggaaagg
     1501 gagctagggc tctggaggct cctggatgcg gaccacggcc cctggccaga gacctggtgg
     1561 atagcgtgga cgatgctgag gggctgtacg tggccgtgga aagatgcccc ctgtgtaata
     1621 ccacacggag aaggctgaca tgcgccaagt gtgtgcagtc cggggatttc gtgtattttg
     1681 acggacgcga tcgggagaga ttcattgaca agaaagaaag gctgtcccgc ctgaagtcta
     1741 aacaggagga atttcagaaa gaggtgctga aggccatgga gggcaagtgg atcactgatc
     1801 agctgaggtg gaaaatcatg tcctgcaaga tgcgcattga gcagctgaaa cagaccatct
     1861 gtaagggaaa cgaggaaatg gagaaaaata gcgaaggcct gctgaagaca aaagaaaaga
     1921 atcagaagct gtactcccgg gctcagagac accaggagaa gaaagaaaaa attcagaggc
     1981 ataaccgcaa gctgggcgat ctggtggaga agaaaaccat cgacctgagg tctcactatg
     2041 aacgcctggc caatctgcgc cggagccata tcctggagct gacatccgtg atcttcccaa
     2101 ttgaggaagt gaaaactgga gtgagggacc ccgctgatgt gagctccgag agtgactcag
     2161 ccatgacctc tagtacagtg agtaagctgg ctgaagccag aaggactacc tacctgtcag
     2221 gccgctgggt gtgcgacgat cacaacggcg atacaagcat ctccattact gggccctgga
     2281 ttagcctgcc taacaatggc gactatagtg cctactattc atgggtggag gaaaagaaaa
     2341 caactcaggg gccagatatg gagcagtcta atcccgctta cactatcagt gccgctctgt
     2401 gctatgccac ccagctggtg aacatcctgt ctcatattct ggacgtgaac ctgcccaaga
     2461 aactgtgcaa ttcagagttc tgtggcgaaa acctgagcaa acagaagttt actagggctg
     2521 tgaagaaact gaacgccaat attctgtacc tgtgttttag ccagcacgtg aatctggatc
     2581 agctgcagcc tctgcatacc ctgcgcaacc tgatgtatct ggtgtcccct tcaagcgagc
     2641 acctgggaag gtctggacca ttcgaagtga gagctgatct ggaggaaagc atggagtttg
     2701 tggaccctgg agtggccggc gagtctgacg aaagtgggga tgagcgggtg tctgacgagg
     2761 aaactgatct gggaaccgac tgggaaaacc tgcctagtcc aagattctgt gacatcccat
     2821 ctcagagtgt ggaggtgtca cagagccagt ccacacaggc ttccccccct attgcttcct
     2881 ctagtgctgg aggaatgatc tcaagcgccg ctgcctcagt gaccagctgg tttaaggcct
     2941 acacaggcca taggtgactc gagactagta tcgcgataat tcactcctca ggtgcaggct
     3001 gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat
     3061 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
     3121 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     3181 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
     3241 agtttggcaa catatgccat atgctggctg ccatgaacaa aggtggctat aaagaggtca
     3301 tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg
     3361 aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt
     3421 ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc
     3481 tgtccctctt ctcttatgaa gatccctcga cctgcagccc aagcttggcg taatcatggt
     3541 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3601 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3661 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc gatccgctgc
     3721 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3781 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3841 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3901 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3961 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4021 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4081 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4141 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4201 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4261 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4321 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4381 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4441 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4501 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4561 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4621 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4681 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4741 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4801 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4861 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4921 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4981 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5041 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5101 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5161 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5221 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5281 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5341 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5401 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5461 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5521 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5581 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5641 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5701 ctggtcgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     5761 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     5821 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     5881 tagggacttt ccattgacgt caatgggtgg actatttacg gtaaactgcc cacttggcag
     5941 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     6001 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     6061 acgtattagt catcgctatt accatgggtc gaggtgagcc ccacgttctg cttcactctc
     6121 cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt
     6181 gcagcgatgg gggcgggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg
     6241 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
     6301 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
     6361 ggcgggagtc gctgcgttgc cttcgccccg tgccccgctc cgcgccgcct cgcgccgccc
     6421 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
     6481 tccgggctgt aattagcgct tggtttaatg acggctcgtt tcttttctgt ggctgcgtga
     6541 aagccttaaa gggctccggg agggcccttt gtgcgggggg gagcggctcg gggggtgcgt
     6601 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggcccgcg ctgcccggcg gctgtgagcg
     6661 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcgtgtgc gcgaggggag cgcggccggg
     6721 ggcggtgccc cgcggtgcgg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     6781 gcgtgggggg gtgagcaggg ggtgtgggcg cggcggtcgg gctgtaaccc ccccctgcac
     6841 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tgcggggcgt
     6901 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     6961 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggccccgg agcgccggcg
     7021 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     7081 agggacttcc tttgtcccaa atctggcgga gccgaaatct gggaggcgcc gccgcacccc
     7141 ctctagcggg cgcgggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     7201 cttcgtgcgt cgccgcgccg ccgtcccctt ctccatctcc agcctcgggg ctgccgcagg
     7261 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     7321 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     7381 gcaacgtgct ggttgttgtg ctgtctcatc attttg
//
