LOCUS       Exported                8364 bp ds-DNA     circular SYN 22-FEB-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8364)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, May 17, 2021 from SnapGene Viewer 5.2.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8364
                     /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..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     misc_feature    1762..5379
                     /label=S ecto dFu 2P
     misc_feature    5389..5469
                     /label=Foldon
     CDS             5476..5499
                     /label=PreScission site
     CDS             5503..5526
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5503..5520
                     /label=6His
     CDS             5506..5523
                     /label=6His
     CDS             5509..5526
                     /label=6His
     CDS             5533..5556
                     /label=Strep tag II
     CDS             5596..5619
                     /label=Strep tag II
     polyA_signal    5786..5841
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6196..6216)
                     /label=M13-rev
     primer_bind     complement(6200..6216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_binding    complement(6222..6244)
                     /label=LacO
     protein_bind    6224..6240
                     /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(6248..6278)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        complement(6249..6278)
                     /label=lac
     protein_bind    6293..6314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6596..7224)
                     /direction=LEFT
                     /label=ColE1 origin
     rep_origin      complement(6614..7202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7373..8233)
                     /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"
     CDS             complement(7376..8035)
                     /label=AmpR
     promoter        complement(8234..8338)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(8275..8303)
                     /label=Amp prom
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 gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gcgcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttgttgt gctgtctcat cattttggca aagaattgcg gccgtctcag
     1741 gccgagttcg gtaccgccac catgttcgtg ttcctggtgc tcctgcctct ggtgagcagc
     1801 cagtgcgtga acctgaccac ccgaacccag ctcccaccag cctacaccaa cagctttaca
     1861 cggggcgtgt actaccctga caaggtgttc agatctagcg tcctgcacag cactcaggac
     1921 ctcttcctgc cgttcttcag caacgtgaca tggttccacg ccatcagcgg cacaaacgga
     1981 accaagcggt ttgataaccc cgtcctgcca ttcaatgatg gagtttactt cgccagtacc
     2041 gagaagagta acatcatccg gggctggatc ttcggcacca ccctggatag caaaacacag
     2101 agcctcctga tcgtgaacaa tgccacgaac gtcgtgatca aggtgtgcga gttccagttt
     2161 tgcaatgatc ctttcctggg tgtgtactac cacaagaaca acaagagctg gatggaaagc
     2221 gagttcagag tctacagcag cgccaacaac tgcacattcg agtacgtctc tcagcctttt
     2281 ctgatggacc ttgaggggaa acaaggcaac ttcaagaacc tgagagaatt cgtgttcaag
     2341 aacatcgacg gctacttcaa aatctactcc aagcacacac ccatcaacct ggtccgggac
     2401 ctccctcagg gcttcagcgc cctggaaccc ctggtcgacc tgcccatagg catcaacata
     2461 acgcggttcc aaaccctgct ggccctgcat agatcctacc tgactcctgg cgacagcagc
     2521 agcggatgga ccgccggagc tgcagcctac tatgtgggct acctgcaacc tagaaccttc
     2581 ctgctgaagt acaacgagaa cggcacaatc acagacgccg tcgactgcgc cctggaccct
     2641 ctctctgaga caaagtgcac cctgaagtcc ttcaccgtgg aaaagggcat ctaccagacc
     2701 agcaacttcc gggtgcagcc tacagagagc atcgtgcgat ttccaaacat taccaacctc
     2761 tgccccttcg gcgaggtgtt taacgccaca agatttgcct ccgtttacgc ctggaataga
     2821 aagagaatca gcaattgtgt ggccgactac tccgtgctgt ataacagcgc ctctttcagc
     2881 accttcaagt gctacggcgt ttccccaaca aagctgaatg acctgtgctt caccaacgtg
     2941 tacgccgact ccttcgtaat tagaggcgat gaggtgcggc agatcgcacc aggccagacc
     3001 ggtaagatcg ctgactacaa ctataagctg cctgatgatt ttacaggctg cgtgatcgcc
     3061 tggaactcta acaacctgga tagcaaggtg ggcggcaact acaactacct gttccggctg
     3121 tttcgcaagt ctaacctgaa acctttcgag agagacatct ccacagagat ctaccaggcc
     3181 ggttctacac cttgtaacgg ggtggaaggc ttcaactgtt acttccctct gcaaagctac
     3241 ggcttccagc ctaccaatgg agtcggctac cagccatacc gggtggtcgt gctgtccttc
     3301 gagttactcc acgcccccgc caccgtctgc ggtcctaaga agtccaccaa tctggttaag
     3361 aacaaatgcg tgaacttcaa cttcaacggc ctgaccggga ccggcgtgct gaccgaaagc
     3421 aacaaaaagt tcctcccctt ccagcagttc ggccgtgata tcgctgacac cacagatgcc
     3481 gtcagagatc cacagaccct ggaaatcctg gatattacac cctgctcctt cggaggagtt
     3541 tctgtgatca cccccgggac caataccagc aaccaggtgg ctgtgctgta ccaaggtgtt
     3601 aactgcaccg aggttcctgt ggccatccac gccgatcagc tgacacctac ttggagagtg
     3661 tactccactg gctccaatgt gttccagacc agggccggat gtctgatcgg cgccgagcac
     3721 gtgaataaca gttacgagtg cgacatccct atcggcgccg gcatctgtgc cagctaccag
     3781 acccagacaa acagccctgg gtctgcttcc tctgtagcta gccagagcgt catcgcctac
     3841 accatgagcc tgggcgcaga gaacagcgtg gcctattcca acaactctat cgccattccc
     3901 accaacttta caattagcgt cacaacagag atcctgcccg tgagcatgac caagaccagc
     3961 gtggactgta caatgtacat ctgtggcgac agcactgaat gcagcaacct gctgctgcaa
     4021 tacggctcct tttgcaccca actgaaccgg gcgctgaccg gaatcgccgt ggaacaggac
     4081 aaaaataccc aggaggtgtt cgcccaagtg aagcagatct acaagacccc acctatcaag
     4141 gacttcggcg gctttaactt tagccagatt ctccctgatc cttctaagcc tagcaagcgg
     4201 agctttatcg aggatctgct gttcaacaag gtcaccctgg ccgatgccgg ctttatcaaa
     4261 cagtatggcg attgcctggg cgacatagcc gccagagatc tgatctgcgc ccagaaattc
     4321 aacggcctga cagttctccc acctctgctg accgacgaga tgatcgctca gtacacctct
     4381 gccctgctgg ctggcaccat cacatctggg tggacatttg gcgccggcgc cgccctgcag
     4441 atcccctttg ccatgcagat ggcctataga ttcaacggaa tcggcgtgac ccagaacgtg
     4501 ctgtatgaaa accagaagct gatcgctaac cagttcaatt ctgccatcgg caagatccag
     4561 gactccctct cctctaccgc cagcgccctg ggcaaactgc aggacgtggt gaatcagaac
     4621 gcccaagccc tgaacaccct ggtgaagcag ctcagcagca attttggcgc catcagctct
     4681 gtgctgaacg atatcctgtc tagactggac aaggtggaag ccgaagtcca gatcgataga
     4741 ctgatcacag gcagactgca gtccctgcaa acctacgtga cccaacagct gatcagggcc
     4801 gctgaaataa gagccagcgc caatctcgcc gctaccaaga tgtccgagtg tgtgctggga
     4861 cagtctaaac gcgttgactt ctgcggcaaa ggctatcacc tgatgagctt cccccagagc
     4921 gcgccgcacg gcgtggtgtt cctgcatgtg acatacgtgc ctgcccaaga gaagaatttc
     4981 acaaccgccc ctgccatctg ccacgacggc aaggcccact tcccaagaga gggcgttttc
     5041 gtttccaatg gcacacactg gttcgtgaca caaagaaact tctacgaacc ccagattatc
     5101 accaccgaca acaccttcgt gagtggcaat tgtgacgtgg tcatcggaat cgtgaacaac
     5161 acagtgtacg accctctgca acctgagctg gactctttta aggaagagct ggacaagtac
     5221 tttaaaaacc acaccagccc cgatgtggac ctgggcgaca tcagtggcat taacgccagc
     5281 gtggtgaaca tccaaaagga aatcgacaga ctgaacgagg tggccaagaa cctgaacgag
     5341 tccctgatcg acctgcagga gctcggcaaa tacgagcagg gatccggata catccccgag
     5401 gcccccagag atggccaggc ctacgtgcgg aaggacggcg agtgggtact gctgagcaca
     5461 ttcctgggca gatccctgga ggtgctgttc cagggcccag gccatcacca ccatcaccac
     5521 catcatagcg cctggtccca cccccagttc gagaagggcg gcggtagtgg agggggcgga
     5581 tctggcggct cagcttggag ccacccccag ttcgaaaagt gataatgact cgagcgataa
     5641 ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     5701 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
     5761 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     5821 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     5881 cagaatgagt atttggttta gagtttggca acatatgcca tatgctggct gccatgaaca
     5941 aaggtggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     6001 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     6061 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     6121 ctgactactc ccagtcatag ctgtccctct tctcttatga agatccctcg acctgcagcc
     6181 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6241 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6301 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6361 gccagcggat cgatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6421 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6481 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6541 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6601 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6661 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6721 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6781 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     6841 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6901 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6961 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7021 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7081 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7141 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7201 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7261 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7321 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7381 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7441 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7501 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7561 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7621 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7681 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7741 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7801 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7861 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7921 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7981 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8041 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8101 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8161 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8221 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8281 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8341 catttccccg aaaagtgcca cctg
//
