LOCUS       Exported                7136 bp DNA     circular SYN 29-APR-2022
DEFINITION  Guo_FM_1_5xST     coverage: 1.16e+03x.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7136)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 2, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      925..1513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    1924..1945
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1960..1990
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1998..2014
                     /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     2022..2038
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             2299..2932
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    2979..3104
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3601..3834
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4019..4063
                     /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    4356..4473
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4516..4914
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     regulatory      4917..4926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4929..4949
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4953..4979
                     /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             4980..4991
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             5397..5453
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5469..5525
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5541..5597
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5613..5669
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5685..5741
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     misc_feature    5824..6412
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6295..6306)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6487..6720
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        6925..7029
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7030..7136,1..754)
                     /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"
ORIGIN
        1 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
       61 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
      121 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
      181 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
      241 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
      301 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
      361 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
      421 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
      481 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
      541 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      601 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      661 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      721 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
      781 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
      841 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
      901 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
      961 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1021 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1081 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1141 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1201 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1261 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1321 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1381 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1441 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1501 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1561 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1621 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     1681 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     1741 ttaatgcagc aagctcatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     1801 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     1861 aaaaagctcc ccgtggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     1921 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     1981 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacat
     2041 gattacgaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2101 actcatcaat gtatcttatc atgtctggat caactggata actcaagcta accaaaatca
     2161 tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt ttcatttact
     2221 ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg
     2281 tactacaaac ttagtagttg gaagggctaa ttcactccca aagaagacaa gatatccttg
     2341 atctgtggat ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc
     2401 caggggtcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag
     2461 ataaggtaga agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc
     2521 atgggatgga tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat
     2581 ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt
     2641 gctacaaggg actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg
     2701 agtggcgagc cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc
     2761 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     2821 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     2881 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     2941 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     3001 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     3061 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     3121 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     3181 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     3241 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     3301 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     3361 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     3421 gtaagaccac cgcacagcaa gcggccggcc gctgatcttc agacctggag gaggagatat
     3481 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     3541 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     3601 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     3661 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     3721 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     3781 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     3841 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     3901 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     3961 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     4021 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     4081 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     4141 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     4201 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     4261 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta
     4321 tcgccgaatt cacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     4381 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     4441 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     4501 ccagtttgga ctagtggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcc
     4561 tcacggcgag cgctgccacg tcagacgaag ggcgcagcga gcgtcctgat ccttccgccc
     4621 ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc
     4681 agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga
     4741 gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg
     4801 gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc
     4861 agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggcggccg
     4921 ccatgggccc aaaaaagaaa agaaaagttg gctaccccta cgacgtgccc gactacgcca
     4981 tcgaaggccg ccatatgcta gcctccaaaa ccatcgttct ttcggtcggc gaggctactc
     5041 gcactctgac tgagatccag tccaccgcag accgtcagat cttcgaagag aaggtcgggc
     5101 ctctggtggg tcggctgcgc ctcacggctt cgctccgtca aaacggagcc aagaccgcgt
     5161 atcgcgtcaa cctaaaactg gatcaggcgg acgtcgttga ttccggactt ccgaaagtgc
     5221 gctacactca ggtatggtcg cacgacgtga caatcgttgc gaatagcacc gaggcctcgc
     5281 gcaaatcgtt gtacgatttg accaagtccc tcgtcgcgac ctcgcaggtc gaagatcttg
     5341 tcgtcaacct tgtgccgctg ggccgtgcgg atccaatcat cggtccgggt tctggagaag
     5401 agctactatc caagaattat catcttgaga acgaggtagc gcgtttgaag aagggttccg
     5461 gctcaggaga ggaactgctc tcgaagaact atcatcttga aaatgaggtc gctcgattaa
     5521 aaaagggatc gggcagtggt gaggaactac tttcaaagaa ttaccacctc gaaaacgaag
     5581 tagctcgatt aaagaaaggt tcagggtcgg gtgaagaatt actgagtaaa aattatcatc
     5641 tggaaaatga ggtagcgaga ctaaaaaagg ggagtggttc tggcgaggaa ttgctatcga
     5701 aaaattatca tcttgagaac gaagttgcta ggctcaaaaa gggctcaggc tcaggcaccg
     5761 cggtaaacgg taccagattc accggtaact atctcgagct gtacaagtaa atcgatagat
     5821 cctaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     5881 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     5941 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     6001 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     6061 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     6121 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     6181 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     6241 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     6301 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     6361 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctgagatc
     6421 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     6481 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     6541 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     6601 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     6661 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     6721 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     6781 agagtgagag gcccgggtta attaaggaaa gggctagatc attcttgaag acgaaagggc
     6841 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     6901 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6961 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     7021 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     7081 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaaga
//
