LOCUS       Exported                4800 bp ds-DNA     circular SYN 28-JUL-2022
DEFINITION  pEH59.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-PE2.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4800)
  TITLE     Liu Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4800)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 29, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4800
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="/Color=#99ff99"
                     /note="/modified_by=jsfg"
                     /note="color: #a6acb3"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /note="color: #ffffff"
     promoter        626..644
                     /label=T7 promoter
                     /note="/created_by=Joung Lab"
                     /note="color: #ffffff"
     regulatory      649..660
                     /label=KOZAK
                     /note="/created_by=jsfg"
                     /note="color: #ffffff"
     misc_feature    658..714
                     /label=bpNLS
                     /label=nonstandard type:  NLS
                     /note="/created_by=jsfg"
                     /note="color: #a6acb3"
     variation       715..1992
                     /label=marathonRT codon optimized
                     /label=nonstandard type:  mutation
                     /note="/created_by=User"
                     /note="color: #a6acb3"
     misc_feature    1993..2004
                     /label=linker
                     /label=nonstandard type:  linker
                     /note="/created_by=jsfg"
                     /note="/modified_by=jsfg"
                     /note="color: #a6acb3"
     misc_feature    2005..2055
                     /label=bpNLS
                     /label=nonstandard type:  NLS
                     /note="/created_by=jsfg"
                     /note="color: #a6acb3"
     regulatory      2056..2058
                     /label=STOP
                     /note="/Color=#000000"
                     /note="/created_by=jsfg"
                     /note="color: #ffffff"
     CDS             2064..2081
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #993366"
                     /translation="HHHHHH"
     polyA_signal    2110..2334
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     protein_bind    2429..2445
                     /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)."
                     /note="color: #31849b"
     promoter        complement(2453..2483)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="color: #ffffff; direction: LEFT"
     protein_bind    2498..2519
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the
                     presenceof cAMP."
                     /note="color: #31849b"
     rep_origin      complement(2807..3395)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="/Color=#99ffff"
                     /note="/modified_by=jsfg"
                     /note="color: #ffff00"
     CDS             complement(3566..4426)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /note="/Color=#ffdd55"
                     /note="/modified_by=jsfg"
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4189..4208
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        complement(4427..4531)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
      661 aaacggacag ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtcgacacc
      721 agcaatctga tggaacagat cctgagcagc gacaacctga accgggccta cctgcaggtg
      781 gtgagaaata aaggcgctga aggcgttgat ggcatgaagt acaccgagct gaaggagcat
      841 ctggccaaga acggcgagac aatcaagggc cagctgagaa ccagaaagta taagcctcag
      901 ccagctagac gggtggaaat ccccaagccc gatggcggag tgcggaacct gggagtgcca
      961 acagtcacag accggttcat ccagcaggct atcgcccaag tgctgacccc tatctacgag
     1021 gaacagtttc acgaccactc ttacggcttc cggcccaaca gatgcgccca gcaagccatc
     1081 ctgacagccc tgaacatcat gaacgatggt aatgactgga tcgtggacat cgacctggaa
     1141 aagtttttcg ataccgtgaa tcacgataag ctgatgacgc tgattggcag aaccatcaag
     1201 gacggcgacg tgatctctat tgtgcgcaag tacctcgtgt ccggcatcat gatcgatgac
     1261 gagtacgaag atagcatcgt gggaacacct cagggcggca acctgtctcc tctgctggcc
     1321 aacatcatgc tgaacgagct ggataaggag atggaaaaaa ggggcctgaa cttcgtgcgg
     1381 tacgccgacg actgcatcat catggtcggc tccgagatga gcgccaacag agtcatgcgg
     1441 aacatcagca gattcatcga agagaagctg ggcctgaaag tgaacatgac caagtccaag
     1501 gtggacagac ctagcggact gaagtacttg ggctttggct tctacttcga ccccagagcc
     1561 caccagttca aggccaagcc tcacgccaag agcgtggcta agttcaaaaa gagaatgaaa
     1621 gagctgacct gtagaagctg gggcgtgtct aacagctaca aggtggaaaa actgaatcaa
     1681 ctgatcagag gctggatcaa ctacttcaag atcggcagca tgaagaccct gtgtaaagag
     1741 ctggacagca gaatcaggta cagactgcgg atgtgcatct ggaagcagtg gaaaacccct
     1801 cagaaccagg agaaaaacct ggtcaagctt ggaattgaca gaaataccgc cagaagagtg
     1861 gcctatacag gcaagcgaat cgcctacgtg tgcaacaagg gcgccgtgaa cgtggctatc
     1921 agcaacaagc ggctggccag cttcggcctg atctctatgc tggactacta catcgagaag
     1981 tgcgtgacct gctctggcgg ctcaaaaaga accgccgacg gcagcgaatt cgagcccaag
     2041 aagaagagga aagtctaacc ggtcatcatc accatcacca ttgagtttaa acccgctgat
     2101 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2161 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag aaaattgcat
     2221 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2281 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     2341 aggcggaaag aaccagctgg ggctcgatac cgtcgacctc tagctagagc ttggcgtaat
     2401 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2461 gagccggaag cataaagtgt aaagcctagg gtgcctaatg agtgagctaa ctcacattaa
     2521 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2581 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2641 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2701 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2761 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2821 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2881 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2941 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3001 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3061 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3121 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3181 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3241 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3301 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3361 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3421 ggtctgacac tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3481 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3541 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3601 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3661 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3721 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3781 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3841 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3901 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3961 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4021 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4081 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4141 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4201 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4261 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4321 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4381 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4441 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4501 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4561 tcgacggatc gggagatcga tctcccgatc ccctagggtc tactctcagt acaatctgct
     4621 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     4681 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     4741 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
