LOCUS       Exported                5553 bp ds-DNA     circular SYN 28-JUL-2022
DEFINITION  pEH45.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-PE2.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5553)
  TITLE     Liu Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5553)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 29, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5553
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     source          715..2745
                     /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"
     CDS             715..2745
                     /label=p80 MMLV-RT - PENTAMUTANT
                     /note="/created_by=jsfg"
                     /note="color: #993366; direction: RIGHT"
     variation       1312..1314
                     /label=D200N
                     /label=nonstandard type:  mutation
                     /note="/created_by=jsfg"
                     /note="color: #a6acb3"
     variation       1630..1632
                     /label=T306K
                     /label=nonstandard type:  mutation
                     /note="/created_by=jsfg"
                     /note="color: #a6acb3"
     variation       1651..1653
                     /label=W313F
                     /label=nonstandard type:  mutation
                     /note="/created_by=jsfg"
                     /note="color: #a6acb3"
     variation       1702..1704
                     /label=T330P
                     /label=nonstandard type:  mutation
                     /note="/created_by=jsfg"
                     /note="color: #a6acb3"
     variation       2521..2523
                     /label=L603W
                     /label=nonstandard type:  mutation
                     /note="/created_by=jsfg"
                     /note="color: #a6acb3"
     misc_feature    2746..2757
                     /label=linker
                     /label=nonstandard type:  linker
                     /note="/created_by=jsfg"
                     /note="/modified_by=jsfg"
                     /note="color: #a6acb3"
     misc_feature    2758..2808
                     /label=bpNLS
                     /label=nonstandard type:  NLS
                     /note="/created_by=jsfg"
                     /note="color: #a6acb3"
     regulatory      2809..2811
                     /label=STOP
                     /note="/Color=#000000"
                     /note="/created_by=jsfg"
                     /note="color: #ffffff"
     CDS             2817..2834
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #993366"
                     /translation="HHHHHH"
     polyA_signal    2863..3087
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     protein_bind    3182..3198
                     /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(3206..3236)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="color: #ffffff; direction: LEFT"
     protein_bind    3251..3272
                     /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(3560..4148)
                     /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(4319..5179)
                     /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     4942..4961
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        complement(5180..5284)
                     /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 agtcacccta
      721 aatatagaag atgagtatcg gctacatgag acctcaaaag agccagatgt ttctctaggg
      781 tccacatggc tgtctgattt tcctcaggcc tgggcggaaa ccgggggcat gggactggca
      841 gttcgccaag ctcctctgat catacctctg aaagcaacct ctacccccgt gtccataaaa
      901 caatacccca tgtcacaaga agccagactg gggatcaagc cccacataca gagactgttg
      961 gaccagggaa tactggtacc ctgccagtcc ccctggaaca cgcccctgct acccgttaag
     1021 aaaccaggga ctaatgatta taggcctgtc caggatctga gagaagtcaa caagcgggtg
     1081 gaagacatcc accccaccgt gcccaaccct tacaacctct tgagcgggct cccaccgtcc
     1141 caccagtggt acactgtgct tgatttaaag gatgcctttt tctgcctgag actccacccc
     1201 accagtcagc ctctcttcgc ctttgagtgg agagatccag agatgggaat ctcaggacaa
     1261 ttgacctgga ccagactccc acagggtttc aaaaacagtc ccaccctgtt taatgaggca
     1321 ctgcacagag acctagcaga cttccggatc cagcacccag acttgatcct gctacagtac
     1381 gtggatgact tactgctggc cgccacttct gagctagact gccaacaagg tactcgggcc
     1441 ctgttacaaa ccctagggaa cctcgggtat cgggcctcgg ccaagaaagc ccaaatttgc
     1501 cagaaacagg tcaagtatct ggggtatctt ctaaaagagg gtcagagatg gctgactgag
     1561 gccagaaaag agactgtgat ggggcagcct actccgaaga cccctcgaca actaagggag
     1621 ttcctaggga aggcaggctt ctgtcgcctc ttcatccctg ggtttgcaga aatggcagcc
     1681 cccctgtacc ctctcaccaa accggggact ctgtttaatt ggggcccaga ccaacaaaag
     1741 gcctatcaag aaatcaagca agctcttcta actgccccag ccctggggtt gccagatttg
     1801 actaagccct ttgaactctt tgtcgacgag aagcagggct acgccaaagg tgtcctaacg
     1861 caaaaactgg gaccttggcg tcggccggtg gcctacctgt ccaaaaagct agacccagta
     1921 gcagctgggt ggcccccttg cctacggatg gtagcagcca ttgccgtact gacaaaggat
     1981 gcaggcaagc taaccatggg acagccacta gtcattctgg ccccccatgc agtagaggca
     2041 ctagtcaaac aaccccccga ccgctggctt tccaacgccc ggatgactca ctatcaggcc
     2101 ttgcttttgg acacggaccg ggtccagttc ggaccggtgg tagccctgaa cccggctacg
     2161 ctgctcccac tgcctgagga agggctgcaa cacaactgcc ttgatatcct ggccgaagcc
     2221 cacggaaccc gacccgacct aacggaccag ccgctcccag acgccgacca cacctggtac
     2281 acggatggaa gcagtctctt acaagaggga cagcgtaagg cgggagctgc ggtgaccacc
     2341 gagaccgagg taatctgggc taaagccctg ccagccggga catccgctca gcgggctgaa
     2401 ctgatagcac tcacccaggc cctaaagatg gcagaaggta agaagctaaa tgtttatact
     2461 gatagccgtt atgcttttgc tactgcccat atccatggag aaatatacag aaggcgtggg
     2521 tggctcacat cagaaggcaa agagatcaaa aataaagacg agatcttggc cctactaaaa
     2581 gccctctttc tgcccaaaag acttagcata atccattgtc caggacatca aaagggacac
     2641 agcgccgagg ctagaggcaa ccggatggct gaccaagcgg cccgaaaggc agccatcaca
     2701 gagactccag acacctctac cctcctcata gaaaattcat caccctctgg cggctcaaaa
     2761 agaaccgccg acggcagcga attcgagccc aagaagaaga ggaaagtcta accggtcatc
     2821 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     2881 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2941 ctgtcctttc ctaataaaat gagaaaattg catcgcattg tctgagtagg tgtcattcta
     3001 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3061 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga
     3121 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     3181 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3241 agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3301 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3361 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3421 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3481 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3541 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3601 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3661 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3721 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3781 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3841 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3901 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3961 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4021 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4081 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4141 gatctcaaga agatcctttg atcttttcta cggggtctga cactcagtgg aacgaaaact
     4201 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4261 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4321 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4381 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4441 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4501 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4561 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4621 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4681 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4741 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4801 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4861 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4921 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4981 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5041 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5101 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5161 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5221 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5281 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cgatctcccg
     5341 atcccctagg gtctactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     5401 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     5461 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     5521 tgcttcgcga tgtacgggcc agatatacgc gtt
//
