LOCUS       Exported                7493 bp DNA     circular SYN 29-APR-2022
DEFINITION  Guo_FM_2_10xST     coverage: 2.88e+03x.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7493)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 2, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(616..632)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    640..656
                     /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)."
     promoter        complement(664..694)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    709..730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1141..1729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1900..2760)
                     /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"
     promoter        complement(2761..2865)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3070..3303
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3378..3966
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3484..3495
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(4055..4111)
                     /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             complement(4127..4183)
                     /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             complement(4199..4255)
                     /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             complement(4271..4327)
                     /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             complement(4343..4399)
                     /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             complement(4415..4471)
                     /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             complement(4487..4543)
                     /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             complement(4559..4615)
                     /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             complement(4631..4687)
                     /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             complement(4703..4759)
                     /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             complement(5156..5167)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(5168..5194)
                     /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             complement(5198..5218)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      5221..5230
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5233..5631)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     misc_feature    5674..5791
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(6084..6128)
                     /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    6313..6546
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    7043..7168
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             join(7215..7493,1..355)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 agtcccttgt agcaagctcg atatcagcag ttcttgaagt actccggatg cagctctcgg
       61 gccacgtgat gaaatgctag gcggctgtca aacctccact ctaacacttc tctctccggg
      121 tcatccatcc catgcaggct cacagggtgt aacaagctgg tgttctctcc tttattggcc
      181 tcttctacct tatctggctc aactggtact agcttgtagc accatccaaa ggtcagtgga
      241 tatctgaccc ctggccctgg tgtgtagttc tgctaatcag ggaagtagcc ttgtgtgtgg
      301 tagatccaca gatcaaggat atcttgtctt ctttgggagt gaattagccc ttccaactac
      361 taagtttgta gtacatattt aacaaataca atttctttaa aatgaaaata attcagagga
      421 atcacaggtt tagagtaaat gaaaccacag gtaattggca gtggtaatag ggtatggggt
      481 gggaagtttg ggatgatttt ggttagcttg agttatccag ttgatccaga catgataaga
      541 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
      601 gaaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
      661 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
      721 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
      781 cggggagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
      841 gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatgag
      901 cttgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
      961 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1021 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1081 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1141 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1201 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1261 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1321 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1381 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1441 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1501 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1561 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1621 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1681 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1741 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1801 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     1861 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     1921 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     1981 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2041 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2101 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2161 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2221 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2281 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     2341 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     2401 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     2461 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg
     2521 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     2581 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     2641 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     2701 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     2761 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     2821 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     2881 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     2941 tatcacgagg ccctttcgtc ttcaagaatg atctagccct ttccttaatt aacccgggcc
     3001 tctcactctc tgatattcat ttctttgcaa gttataaata ctgaataata agatgacatg
     3061 aactactact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     3121 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     3181 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     3241 agagagaccc agtacaagca aaaagcagat cttgtcttcg ttgggagtga attagccctt
     3301 ccagtccccc cttttctttt aaaaagtggc taagatctac agctgccttg taagtcattg
     3361 gtcttaaagg atctcaggcg gggaggcggc ccaaagggag atccgactcg tctgagggcg
     3421 aaggcgaaga cgcggaagag gccgcagagc cggcagcagg ccgcgggaag gaaggtccgc
     3481 tggattgagg gccgaaggga cgtagcagaa ggacgtcccg cgcagaatcc aggtggcaac
     3541 acaggcgagc agccaaggaa aggacgatga tttccccgac aacaccacgg aattgtcagt
     3601 gcccaacagc cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt
     3661 ggcaataggg agggggaaag cgaaagtccc ggaaaggagc tgacaggtgg tggcaatgcc
     3721 ccaaccagtg ggggttgcgt cagcaaacac agtgcacacc acgccacgtt gcctgacaac
     3781 gggccacaac tcctcataaa gagacagcaa ccaggattta tacaaggagg agaaaatgaa
     3841 agccatacgg gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt
     3901 aaaaggagca acatagttaa gaataccagt caatctttca caaattttgt aatccagagg
     3961 ttgattagga tctatcgatt tacttgtaca gctcgagata gttaccggtg aatctggtac
     4021 cccatatgtt taccgcggtg cctgagcctg agcccttttt gagcctagca acttcgttct
     4081 caagatgata atttttcgat agcaattcct cgccagaacc actccccttt tttagtctcg
     4141 ctacctcatt ttccagatga taatttttac tcagtaattc ttcacccgac cctgaacctt
     4201 tctttaatcg agctacttcg ttttcgaggt ggtaattctt tgaaagtagt tcctcaccac
     4261 tgcccgatcc cttttttaat cgagcgacct cattttcaag atgatagttc ttcgagagca
     4321 gttcctctcc tgagccggaa cccttcttca aacgcgctac ctcgttctca agatgataat
     4381 tcttggatag tagctcttca ccactgccac ttcccttttt taagcgtgcc acctcatttt
     4441 cgaggtggta gtttttagat aatagctctt ctccagaacc actgcccttt tttagccgag
     4501 ccacttcgtt ttcgagatga taatttttcg agagtaactc ctcacccgat ccggaccctt
     4561 tctttagtcg tgctacttcg ttctccaaat gataattttt gctcagcaac tcctccccac
     4621 ttccgctccc ctttttcagt ctagctacct cattttccag gtggtaattc tttgaaagca
     4681 gttcttctcc actgccagaa cctttcttaa gacgagccac ttcgttctca agatgataat
     4741 tcttgctcaa aagttcttcc ggaccgcttt gtggatccgc acggcccagc ggcacaaggt
     4801 tgacgacaag atcttcgacc tgcgaggtcg cgacgaggga cttggtcaaa tcgtacaacg
     4861 atttgcgcga ggcctcggtg ctattcgcaa cgattgtcac gtcgtgcgac catacctgag
     4921 tgtagcgcac tttcggaagt ccggaatcaa cgacgtccgc ctgatccagt tttaggttga
     4981 cgcgatacgc ggtcttggct ccgttttgac ggagcgaagc cgtgaggcgc agccgaccca
     5041 ccagaggccc gaccttctct tcgaagatct gacggtctgc ggtggactgg atctcagtca
     5101 gagtgcgagt agcctcgccg accgaaagaa cgatggtttt ggaggctagc atatggcggc
     5161 cttcgatggc gtagtcgggc acgtcgtagg ggtagccaac ttttcttttc ttttttgggc
     5221 ccatggcggc cgccaagtga cgatcacagc gatccacaaa caagaaccgc gacccaaatc
     5281 ccggctgcga cggaactagc tgtgccacac ccggcgcgtc cttatataat catcggcgtt
     5341 caccgcccca cggagatccc tccgcagaat cgccgagaag ggactacttt tcctcgcctg
     5401 ttccgctctc tggaaagaaa accagtgccc tagagtcacc caagtcccgt cctaaaatgt
     5461 ccttctgctg atactggggt tctaaggccg agtcttatga gcagcgggcc gctgtcctga
     5521 gcgtccgggc ggaaggatca ggacgctcgc tgcgcccttc gtctgacgtg gcagcgctcg
     5581 ccgtgaggag gggggcgccc gcgggaggcg ccaaaacccg gcgcggaggc cactagtcca
     5641 aactggatct ctgctgtccc tgtaataaac ccgaaaattt tgaatttttg taatttgttt
     5701 ttgtaattct ttagtttgta tgtctgttgc tattatgtct actattcttt cccctgcact
     5761 gtacccccca atcccccctt ttcttttaaa attgtggatg aatactgcca tttgtgaatt
     5821 cggcgatacc gtcgagatcc gttcactaat cgaatggatc tgtctctgtc tctctctcca
     5881 ccttcttctt ctattccttc gggcctgtcg ggtcccctcg gggttgggag gtgggtctga
     5941 aacgataatg gtgaatatcc ctgcctaact ctattcacta tagaaagtac agcaaaaact
     6001 attcttaaac ctaccaagcc tcctactatc attatgaata attttatata ccacagccaa
     6061 tttgttatgt taaaccaatt ccacaaactt gcccatttat ctaattccaa taattcttgt
     6121 tcattctttt cttgctggtt ttgcgattct tcaattaagg agtgtattaa gcttgtgtaa
     6181 ttgttaattt ctctgtccca ctccatccag gtcgtgtgat tccaaatctg ttccagagat
     6241 ttattactcc aactagcatt ccaaggcaca gcagtggtgc aaatgagttt tccagagcaa
     6301 ccccaaatcc ccaggagctg ttgatccttt aggtatcttt ccacagccag gattcttgcc
     6361 tggagctgct tgatgcccca gactgtgagt tgcaacagat gctgttgcgc ctcaatagcc
     6421 ctcagcaaat tgttctgctg ctgcactata ccagacaata attgtctggc ctgtaccgtc
     6481 agcgtcattg acgctgcgcc catagtgctt cctgctgctc ccaagaaccc aaggaacaaa
     6541 gctcctattc ccactgctct tttttctctc tgcaccactc ttctctttgc cttggtgggt
     6601 gctactccta atggttcaat ttttactact ttatatttat ataattcact tctccaattg
     6661 tccctcatat ctcctcctcc aggtctgaag atcagcggcc ggccgcttgc tgtgcggtgg
     6721 tcttactttt gttttgctct tcctctatct tgtctaaagc ttccttggtg tcttttatct
     6781 ctatcctttg atgcacacaa tagagggttg ctactgtatt atataatgat ctaagttctt
     6841 ctgatcctgt ctgaagggat ggttgtagct gtcccagtat ttgtctacag ccttctgatg
     6901 tttctaacag gccaggatta actgcgaatc gttctagctc cctgcttgcc catactatat
     6961 gttttaattt atattttttc tttccccctg gccttaaccg aattttttcc catcgcgatc
     7021 taattctccc ccgcttaata ctgacgctct cgcacccatc tctctccttc tagcctccgc
     7081 tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc gcctcttgcc gtgcgcgctt
     7141 cagcaagccg agtcctgcgt cgagagagct cctctggttt ccctttcgct ttcaagtccc
     7201 tgttcgggcg ccactgctag agattttcca cactgactaa aagggtctga gggatctcta
     7261 gttaccagag tcacacaaca gacgggcaca cactacttga agcactcaag gcaagcttta
     7321 ttgaggctta agcagtgggt tccctagtta gccagagagc tcccaggctc agatctggtc
     7381 taaccagaga gacccagtac aggcaaaaag cagctgctta tatgcaggat ctgagggctc
     7441 gccactcccc agtcccgccc aggccacgcc tccctggaaa gtccccagcg gaa
//
