LOCUS       sequence-481851-        6356 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  E.HA.FLAG Flow Code Plasmid .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6356)
  TITLE     Flow Code Plasmids
REFERENCE   2  (bases 1 to 6356)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pMIHP-10519 E.HA.FLAG
FEATURES             Location/Qualifiers
     source          1..6356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..27
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     155..174
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             374..890
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    954..1295
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1362..1778
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1708..1730
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1746..1762
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1848..2397
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1992..2009)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2219..2238
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      2808..2817
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2877..2903
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2922..2945
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     LTR             3124..3638
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3800..3816
                     /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(join(3824..3830,3831..3848,3849..3854))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3869..3890
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4007..4024)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4178..4766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4258..4277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4937..5728,5729..5797))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5560..5579
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5798..5902)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5970..5988
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6026..6048)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6148..6167
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa tggtgcatgc aaggagatgg
       61 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
      121 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
      181 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggcgattag
      241 tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc
      301 accagctgaa gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc
      361 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
      421 cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg
      481 aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
      541 ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag
      601 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
      661 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
      721 aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta
      781 cccgtattcc caataaagcc tcttgctgtt tgcatccgaa tcgtggactc gctgatcctt
      841 gggagggtct cctcagattg attgactgcc cacctcgggg gtctttcatt tggaggttcc
      901 accgagattt ggagacccct gcccagggac caccgacccc cccgccggga ggtaagctgg
      961 ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc atctaatgtt
     1021 tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
     1081 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
     1141 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
     1201 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
     1261 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
     1321 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
     1381 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1441 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1501 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1561 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1621 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1681 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1741 cctcccttta tccagccctc actccttctc taggcgccgg aattccgaac agtcttccgt
     1801 aatagaagac acaaggctcg agcgggatca attccgcccc ccccctaacg ttactggccg
     1861 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     1921 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     1981 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     2041 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     2101 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     2161 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     2221 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     2281 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     2341 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gataatacca
     2401 tgccagagcc ttctaagtct gcaccagccc ctaagaaggg atctaagaaa gccatctcta
     2461 aggcgcagaa gaaggatggc aagaagcgca agcgcagccg caaggagagc tattcggtgt
     2521 acgtgtacaa ggtgctgaag caagtgcacc ccgacaccgg catctcctcc aaggccatgg
     2581 gcatcatgaa ctcgttcgtg aacgacatct tcgagcgcat cgcgagcgag gcgtcccgcc
     2641 tggcgcatta caacaagcgc tcgaccatca cgtcccggga gatccagacg gccgtgcgcc
     2701 tgctgctgcc cggggagctg gccaagcacg ccgtgtcgga gggcactaag gccgtcacca
     2761 agtacaccag ctccaaggac ccaccggtcg ccaccggatc cacggtcgcc accatgggcg
     2821 gggcacctgt gccctatcca gatcccctgg agcctagggg aggaggcggg ggagggtacc
     2881 catacgatgt gccagattac gctggaggag gcgggggagg ggactacaaa gacgatgacg
     2941 acaagggcat gctgatggtg taagcggccg cgactctaga gtcgacctgc aggcatgcaa
     3001 gcttcaggta gccggctaac gttaacaacc ggtacctcta gaactatagc tagcatgcgc
     3061 aaatttaaag cgctgatatc gataaaataa aagattttat ttagtctcca gaaaaagggg
     3121 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     3181 gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga acagagagac
     3241 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     3301 aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat
     3361 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     3421 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     3481 acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc
     3541 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     3601 tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt cttgaagttg
     3661 gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca attagccact
     3721 gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag cgcagaaaga
     3781 gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3841 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3901 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3961 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4021 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4081 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4141 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4201 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4261 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4321 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4381 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4441 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4501 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4561 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4621 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4681 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4741 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4801 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4861 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4921 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4981 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5041 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5101 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5161 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5221 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5281 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5341 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5401 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5461 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5521 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5581 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5641 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5701 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5761 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5821 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5881 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     5941 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     6001 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     6061 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     6121 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     6181 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     6241 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     6301 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccaggg
//
