LOCUS       Exported                8765 bp ds-DNA     circular SYN 01-DEC-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pHAGE-EFS-MCP-3XBFPnls
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8765)
  AUTHORS   wangyang
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 13, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8765
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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."
     primer_bind     1948..1972
                     /label=5'seq for 3xFP
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2265..2476
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2521..2530
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2527..2877
                     /codon_start=1
                     /label=MCP
                     /translation="MASNFTQFVLVDNGGTGDVTVAPSNFANGIAEWISSNSRSQAYKV
                     TCSVRQSSAQNRKYTIKVEVPKGAWRSYLNMELTIPIFATNSDCELIVKAMQGLLKDGN
                     PIPSAIAANSGIY"
     primer_bind     2890..2910
                     /label=13a_3xBFP_F
     primer_bind     2890..2910
                     /label=13b_3xBFP_F
     CDS             2911..3621
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             3640..4350
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             4369..5079
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             5098..5118
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5125..5145
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    5155..5743
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5626..5637)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5711..5730)
                     /label=3'seq for 3xFP
     LTR             5818..6051
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        6256..6360
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6361..7221
                     /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"
     rep_origin      7392..7980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8391..8412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8427..8457
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8465..8481
                     /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     8489..8505
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 ttgacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcgat
     2221 gctagctgat gcgggccctt cgagtggctc cggtgcccgt cagtgggcag agcgcacatc
     2281 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
     2341 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     2401 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     2461 tgccgccaga acacaggtgt cgtgacgcgg tccggttaag gtgaacgcgt gcaggctggc
     2521 gccaccatgg cttctaactt tactcagttc gttctcgtcg acaatggcgg aactggcgac
     2581 gtgactgtcg ccccaagcaa cttcgctaac gggatcgctg aatggatcag ctctaactcg
     2641 cgttcacagg cttacaaagt aacctgtagc gttcgtcaga gctctgcgca gaatcgcaaa
     2701 tacaccatca aagtcgaggt gcctaaaggc gcctggcgtt cgtacttaaa tatggaacta
     2761 accattccaa ttttcgccac gaattccgac tgcgagctta ttgttaaggc aatgcaaggt
     2821 ctcctaaaag atggaaaccc gattccctca gcaatcgcag caaactccgg catctacggg
     2881 ggatctggta gcggcgctgg atccggaagt atggtgtcta agggcgaaga gctgattaag
     2941 gagaacatgc acatgaagct gtacatggag ggcaccgtgg acaaccatca cttcaagtgc
     3001 acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc
     3061 gagggcggcc ctctcccctt cgccttcgac atcctggcta ctagcttcct ctacggcagc
     3121 aagaccttca tcaaccacac ccagggcatc cccgacttct tcaagcagtc cttccctgag
     3181 ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag
     3241 gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc
     3301 acatccaacg gccctgtgat gcagaagaaa acactcggct gggaggcctt caccgagacg
     3361 ctgtaccccg ctgacggcgg cctggaaggc agaaacgaca tggccctgaa gctcgtgggc
     3421 gggagccatc tgatcgcaaa cgccaagacc acatatagat ccaagaaacc cgctaagaac
     3481 ctcaagatgc ctggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccaac
     3541 aacgaaacct acgtcgagca gcacgaggtg gcagtggcca gatactgcga cctccctagc
     3601 aaactggggc acaagcttaa ttctggcggc cgctctggca tggtgtctaa gggcgaagag
     3661 ctgattaagg agaacatgca catgaagctg tacatggagg gcaccgtgga caaccatcac
     3721 ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc
     3781 aaggtggtcg agggcggccc tctccccttc gccttcgaca tcctggctac tagcttcctc
     3841 tacggcagca agaccttcat caaccacacc cagggcatcc ccgacttctt caagcagtcc
     3901 ttccctgagg gcttcacatg ggagagagtc accacatacg aagacggggg cgtgctgacc
     3961 gctacccagg acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg
     4021 gtgaacttca catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccttc
     4081 accgagacgc tgtaccccgc tgacggcggc ctggaaggca gaaacgacat ggccctgaag
     4141 ctcgtgggcg ggagccatct gatcgcaaac gccaagacca catatagatc caagaaaccc
     4201 gctaagaacc tcaagatgcc tggcgtctac tatgtggact acagactgga aagaatcaag
     4261 gaggccaaca acgaaaccta cgtcgagcag cacgaggtgg cagtggccag atactgcgac
     4321 ctccctagca aactggggca caagcttaat ggtagcatcg atggtagcat ggtgtctaag
     4381 ggcgaagagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtggac
     4441 aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
     4501 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact
     4561 agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc
     4621 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc
     4681 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
     4741 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
     4801 gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
     4861 gccctgaagc tcgtgggcgg gagccatctg atcgcaaacg ccaagaccac atatagatcc
     4921 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
     4981 agaatcaagg aggccaacaa cgaaacctac gtcgagcagc acgaggtggc agtggccaga
     5041 tactgcgacc tccctagcaa actggggcac aagcttaatc tcgagggcag cggctccccc
     5101 aagaaaaaac gcaaggtgga agatcctaag aaaaagcgga aagtgtaact agataatcaa
     5161 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     5221 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     5281 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     5341 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     5401 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     5461 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     5521 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     5581 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     5641 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     5701 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctgagat cctttaagac
     5761 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     5821 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     5881 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     5941 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6001 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     6061 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     6121 ggcccgggtt aattaaggaa agggctagat cattcttgaa gacgaaaggg cctcgtgata
     6181 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     6241 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6301 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6361 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6421 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6481 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6541 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6601 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6661 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6721 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6781 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6841 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6901 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6961 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7021 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     7081 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     7141 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7201 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     7261 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7321 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7381 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7441 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7501 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     7561 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7621 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7681 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7741 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7801 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7861 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7921 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7981 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8041 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     8101 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     8161 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagcaag
     8221 ctcatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     8281 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctccccg
     8341 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     8401 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     8461 ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat tacgaatttc
     8521 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     8581 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
     8641 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
     8701 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
     8761 gtagt
//
