LOCUS       Exported                9097 bp ds-DNA     circular SYN 04-FEB-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9097)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Feb 5, 2020 from SnapGene Viewer 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9097
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     CDS             5939..5959
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5966..6031
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    6186..6241
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6600..6616)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6624..6640
                     /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(6648..6678)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6693..6714
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6773..6968
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6819..6954
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6974..7108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(7347..7935)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8106..8966)
                     /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(8967..9071)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attctgcagt cgacggtacc
     1741 gcgggcccgg gatccaccgg tcgccaccat gaccaagccc tacagcatcg gcctggacat
     1801 cggcaccaat agcgtgggct gggccgtgac caccgacaac tacaaggtgc ccagcaagaa
     1861 aatgaaggtg ctgggcaaca cctccaagaa gtacatcaag aaaaacctgc tgggcgtgct
     1921 gctgttcgac agcggcatta cagccgaggg cagacggctg aagagaaccg ccagacggcg
     1981 gtacacccgg cggagaaaca gaatcctgta tctgcaagag atcttcagca ccgagatggc
     2041 taccctggac gacgccttct tccagcggct ggacgacagc ttcctggtgc ccgacgacaa
     2101 gcgggacagc aagtacccca tcttcggcaa cctggtggaa gagaaggcct accacgacga
     2161 gttccccacc atctaccacc tgagaaagta cctggccgac agcaccaaga aggccgacct
     2221 gagactggtg tatctggccc tggcccacat gatcaagtac cggggccact tcctgatcga
     2281 gggcgagttc aacagcaaga acaacgacat ccagaagaac ttccaggact tcctggacac
     2341 ctacaacgcc atcttcgaga gcgacctgtc cctggaaaac agcaagcagc tggaagagat
     2401 cgtgaaggac aagatcagca agctggaaaa gaaggaccgc atcctgaagc tgttccccgg
     2461 cgagaagaac agcggaatct tcagcgagtt tctgaagctg atcgtgggca accaggccga
     2521 cttcagaaag tgcttcaacc tggacgagaa agccagcctg cacttcagca aagagagcta
     2581 cgacgaggac ctggaaaccc tgctgggata tatcggcgac gactacagcg acgtgttcct
     2641 gaaggccaag aagctgtacg acgctatcct gctgagcggc ttcctgaccg tgaccgacaa
     2701 cgagacagag gccccactga gcagcgccat gattaagcgg tacaacgagc acaaagagga
     2761 tctggctctg ctgaaagagt acatccggaa catcagcctg aaaacctaca atgaggtgtt
     2821 caaggacgac accaagaacg gctacgccgg ctacatcgac ggcaagacca accaggaaga
     2881 tttctatgtg tacctgaaga agctgctggc cgagttcgag ggggccgact actttctgga
     2941 aaaaatcgac cgcgaggatt tcctgcggaa gcagcggacc ttcgacaacg gcagcatccc
     3001 ctaccagatc catctgcagg aaatgcgggc catcctggac aagcaggcca agttctaccc
     3061 attcctggcc aagaacaaag agcggatcga gaagatcctg accttccgca tcccttacta
     3121 cgtgggcccc ctggccagag gcaacagcga ttttgcctgg tccatccgga agcgcaatga
     3181 gaagatcacc ccctggaact tcgaggacgt gatcgacaaa gagtccagcg ccgaggcctt
     3241 catcaaccgg atgaccagct tcgacctgta cctgcccgag gaaaaggtgc tgcccaagca
     3301 cagcctgctg tacgagacat tcaatgtgta taacgagctg accaaagtgc ggtttatcgc
     3361 cgagtctatg cgggactacc agttcctgga ctccaagcag aaaaaggaca tcgtgcggct
     3421 gtacttcaag gacaagcgga aagtgaccga taaggacatc atcgagtacc tgcacgccat
     3481 ctacggctac gatggcatcg agctgaaggg catcgagaag cagttcaact ccagcctgag
     3541 cacataccac gacctgctga acattatcaa cgacaaagaa tttctggacg actccagcaa
     3601 cgaggccatc atcgaagaga tcatccacac cctgaccatc tttgaggacc gcgagatgat
     3661 caagcagcgg ctgagcaagt tcgagaacat cttcgacaag agcgtgctga aaaagctgag
     3721 cagacggcac tacaccggct ggggcaagct gagcgccaag ctgatcaacg gcatccggga
     3781 cgagaagtcc ggcaacacaa tcctggacta cctgatcgac gacggcatca gcaaccggaa
     3841 cttcatgcag ctgatccacg acgacgccct gagcttcaag aagaagatcc agaaggccca
     3901 gatcatcggg gacgaggaca agggcaacat caaagaagtc gtgaagtccc tgcccggcag
     3961 ccccgccatc aagaagggaa tcctgcagag catcaagatc gtggacgagc tcgtgaaagt
     4021 gatgggcggc agaaagcccg agagcatcgt ggtggaaatg gctagagaga accagtacac
     4081 caatcagggc aagagcaaca gccagcagag actgaagaga ctggaaaagt ccctgaaaga
     4141 gctgggcagc aagattctga aagagaatat ccctgccaag ctgtccaaga tcgacaacaa
     4201 cgccctgcag aacgaccggc tgtacctgta ctacctgcag aatggcaagg acatgtatac
     4261 aggcgacgac ctggatatcg accgcctgag caactacgac atcgaccata ttatccccca
     4321 ggccttcctg aaagacaaca gcattgacaa caaagtgctg gtgtcctccg ccagcaaccg
     4381 cggcaagtcc gatgatgtgc ccagcctgga agtcgtgaaa aagagaaaga ccttctggta
     4441 tcagctgctg aaaagcaagc tgattagcca gaggaagttc gacaacctga ccaaggccga
     4501 gagaggcggc ctgagccctg aagataaggc cggcttcatc cagagacagc tggtggaaac
     4561 ccggcagatc accaagcacg tggccagact gctggatgag aagtttaaca acaagaagga
     4621 cgagaacaac cgggccgtgc ggaccgtgaa gatcatcacc ctgaagtcca ccctggtgtc
     4681 ccagttccgg aaggacttcg agctgtataa agtgcgcgag atcaatgact ttcaccacgc
     4741 ccacgacgcc tacctgaatg ccgtggtggc ttccgccctg ctgaagaagt accctaagct
     4801 ggaacccgag ttcgtgtacg gcgactaccc caagtacaac tccttcagag agcggaagtc
     4861 cgccaccgag aaggtgtact tctactccaa catcatgaat atctttaaga agtccatctc
     4921 cctggccgat ggcagagtga tcgagcggcc cctgatcgaa gtgaacgaag agacaggcga
     4981 gagcgtgtgg aacaaagaaa gcgacctggc caccgtgcgg cgggtgctga gttatcctca
     5041 agtgaatgtc gtgaagaagg tggaagaaca gaaccacggc ctggatcggg gcaagcccaa
     5101 gggcctgttc aacgccaacc tgtccagcaa gcctaagccc aactccaacg agaatctcgt
     5161 gggggccaaa gagtacctgg accctaagaa gtacggcgga tacgccggca tctccaatag
     5221 cttcaccgtg ctcgtgaagg gcacaatcga gaagggcgct aagaaaaaga tcacaaacgt
     5281 gctggaattt caggggatct ctatcctgga ccggatcaac taccggaagg ataagctgaa
     5341 ctttctgctg gaaaaaggct acaaggacat tgagctgatt atcgagctgc ctaagtactc
     5401 cctgttcgaa ctgagcgacg gctccagacg gatgctggcc tccatcctgt ccaccaacaa
     5461 caagcggggc gagatccaca agggaaacca gatcttcctg agccagaaat ttgtgaaact
     5521 gctgtaccac gccaagcgga tctccaacac catcaatgag aaccaccgga aatacgtgga
     5581 aaaccacaag aaagagtttg aggaactgtt ctactacatc ctggagttca acgagaacta
     5641 tgtgggagcc aagaagaacg gcaaactgct gaactccgcc ttccagagct ggcagaacca
     5701 cagcatcgac gagctgtgca gctccttcat cggccctacc ggcagcgagc ggaagggact
     5761 gtttgagctg acctccagag gctctgccgc cgactttgag ttcctgggag tgaagatccc
     5821 ccggtacaga gactacaccc cctctagtct gctgaaggac gccaccctga tccaccagag
     5881 cgtgaccggc ctgtacgaaa cccggatcga cctggctaag ctgggcgagg gaggatcccc
     5941 caagaagaag aggaaagtct cgagcgacta caaagaccat gacggtgatt ataaagatca
     6001 tgacatcgat tacaaggatg acgatgacaa gtaaagcggc cgcactcctc aggtgcaggc
     6061 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     6121 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     6181 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     6241 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     6301 gagtttggca acatatgcca tatgctggct gccatgaaca aaggtggcta taaagaggtc
     6361 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     6421 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     6481 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     6541 ctgtccctct tctcttatga agatccctcg acctgcagcc caagcttggc gtaatcatgg
     6601 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6661 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6721 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     6781 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6841 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6901 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6961 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     7021 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     7081 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     7141 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7201 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7261 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7321 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7381 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7441 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7501 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     7561 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7621 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7681 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7741 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     7801 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7861 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7921 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7981 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8041 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8101 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8161 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8221 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8281 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8341 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8401 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8461 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8521 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8581 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8641 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8701 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8761 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8821 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8881 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8941 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     9001 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     9061 ggggttccgc gcacatttcc ccgaaaagtg ccacctg
//
