LOCUS       sequence-108561-        7943 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Used to label reporter mRNAs. Lentiviral expression of the MCP 
            protein fused to HALO..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7943)
  AUTHORS   Halstead JM, Lionnet T, Wilbertz JH, Wippich F, Ephrussi A, Singer 
            RH, Chao JA
  TITLE     Translation. An RNA biosensor for imaging the first round of 
            translation from single cells to living animals.
  JOURNAL   Science. 2015 Mar 20;347(6228):1367-671. doi: 
            10.1126/science.aaa3380.
  PUBMED    25792328
REFERENCE   2  (bases 1 to 7943)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2015-03-20- 20"; volume: "347"; 
            issue: "6228"; pages: "1367-671"
COMMENT     Sequence Label: phage ubc nls ha 2xmcp HALO
FEATURES             Location/Qualifiers
     source          1..7943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2218..2616
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             2631..2651
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2655..2681
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2655..2681
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2682..2693
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3429..4313
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     misc_feature    4333..4921
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4386..4406)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4804..4815)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4996..5229
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5348..5366)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5434..5538
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5539..5607,5608..6399)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5757..5776)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6570..7158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7059..7078
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7312..7329
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7569..7590
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7605..7610,7611..7628,7629..7635)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7643..7659
                     /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)."
     primer_bind     7648..7670
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7667..7683
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7667..7683
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7733..7752
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtggc
     2221 ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cctcacggcg agcgctgcca
     2281 cgtcagacga agggcgcagc gagcgtcctg atccttccgc ccggacgctc aggacagcgg
     2341 cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac
     2401 gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa
     2461 agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat
     2521 tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc
     2581 ggttcttgtt tgtggatcgc tgtgatcgtc acttggcggc cgcaatgggc ccaaaaaaga
     2641 aaagaaaagt tggctacccc tacgacgtgc ccgactacgc catcgaaggc cgccatatgc
     2701 tagccgttaa aatggcttct aactttactc agttcgttct cgtcgacaat ggcggaactg
     2761 gcgacgtgac tgtcgcccca agcaacttcg ctaacgggat cgctgaatgg atcagctcta
     2821 actcgcgttc acaggcttac aaagtaacct gtagcgttcg tcagagctct gcgcagaatc
     2881 gcaaatacac catcaaagtc gaggtgccta aaggcgcctg gcgttcgtac ttaaatatgg
     2941 aactaaccat tccaattttc gccacgaatt ccgactgcga gcttattgtt aaggcaatgc
     3001 aaggtctcct aaaagatgga aacccgattc cctcagcaat cgcagcaaac tccggcatct
     3061 acgccatggc ttctaacttt actcagttcg ttctcgtcga caatggcgga actggcgacg
     3121 tgactgtcgc cccaagcaac ttcgctaacg ggatcgctga atggatcagc tctaactcgc
     3181 gttcacaggc ttacaaagta acctgtagcg ttcgtcagag ctctgcgcag aatcgcaaat
     3241 acaccatcaa agtcgaggtg cctaaaggcg cctggcgttc gtacttaaat atggaactaa
     3301 ccattccaat tttcgccacg aattccgact gcgagcttat tgttaaggca atgcaaggtc
     3361 tcctaaaaga tggaaacccg attccctcag caatcgcagc aaactccggc atctacgcgg
     3421 attctagaga aatcggtact ggctttccat tcgaccccca ttatgtggaa gtcctgggcg
     3481 agcgcatgca ctacgtcgat gttggtccgc gcgatggcac ccctgtgctg ttcctgcacg
     3541 gtaacccgac ctcctcctac gtgtggcgca acatcatccc gcatgttgca ccgacccatc
     3601 gctgcattgc tccagacctg atcggtatgg gcaaatccga caaaccagac ctgggttatt
     3661 tcttcgacga ccacgtccgc ttcatggatg ccttcatcga agccctgggt ctggaagagg
     3721 tcgtcctggt cattcacgac tggggctccg ctctgggttt ccactgggcc aagcgcaatc
     3781 cagagcgcgt caaaggtatt gcatttatgg agttcatccg ccctatcccg acctgggacg
     3841 aatggccaga atttgcccgc gagaccttcc aggccttccg caccaccgac gtcggccgca
     3901 agctgatcat cgatcagaac gtttttatcg agggtacgct gccgatgggt gtcgtccgcc
     3961 cgctgactga agtcgagatg gaccattacc gcgagccgtt cctgaatcct gttgaccgcg
     4021 agccactgtg gcgcttccca aacgagctgc caatcgccgg tgagccagcg aacatcgtcg
     4081 cgctggtcga agaatacatg gactggctgc accagtcccc tgtcccgaag ctgctgttct
     4141 ggggcacccc aggcgttctg atcccaccgg ccgaagccgc tcgcctggcc aaaagcctgc
     4201 ctaactgcaa ggctgtggac atcggcccgg gtctgaatct gctgcaagaa gacaacccgg
     4261 acctgatcgg cagcgagatc gcgcgctggc tgtctactct ggagatttcc ggttaatgaa
     4321 tcgatagatc ctaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     4381 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     4441 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     4501 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     4561 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     4621 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     4681 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     4741 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4801 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4861 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4921 cctgagatcc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     4981 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
     5041 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     5101 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     5161 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     5221 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     5281 gaatatcaga gagtgagagg cccgggttaa ttaaggaaag ggctagatca ttcttgaaga
     5341 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5401 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5461 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5521 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5581 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5641 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5701 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5761 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     5821 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5881 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5941 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6001 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6061 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6121 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6181 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6241 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6301 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6361 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6421 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6481 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6541 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6601 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6661 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6721 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6781 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6841 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6901 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6961 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7021 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7081 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7141 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7201 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7261 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7321 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     7381 attcattaat gcagcaagct catggctgac taattttttt tatttatgca gaggccgagg
     7441 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     7501 tttgcaaaaa gctccccgtg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     7561 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     7621 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     7681 gacatgatta cgaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     7741 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa
     7801 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat
     7861 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa
     7921 atatgtacta caaacttagt agt
//
