LOCUS       sequence-481830-        6386 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  HAT.FLAG.E Flow Code Plasmid .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6386)
  TITLE     Flow Code Plasmids
REFERENCE   2  (bases 1 to 6386)
  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-12524 HAT.FLAG.E
FEATURES             Location/Qualifiers
     source          1..6386
                     /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             2895..2918
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     LTR             3154..3668
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3830..3846
                     /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(3854..3860,3861..3878,3879..3884))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3899..3920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4037..4054)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4208..4796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4288..4307)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4967..5758,5759..5827))
                     /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     5590..5609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5828..5932)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6000..6018
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6056..6078)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6178..6197
                     /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 accatgggca
     2821 aggaccacct gatccacaac gtgcacaagg agttccacgc ccacgcccac aacaagggag
     2881 gaggcggggg aggggactac aaagacgatg acgacaaggg aggaggcggg ggaggggggg
     2941 cacctgtgcc ctatccagat cccctggagc ctaggggcat gctgatggtg taagcggccg
     3001 cgactctaga gtcgacctgc aggcatgcaa gcttcaggta gccggctaac gttaacaacc
     3061 ggtacctcta gaactatagc tagcatgcgc aaatttaaag cgctgatatc gataaaataa
     3121 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     3181 aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga
     3241 agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt
     3301 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg
     3361 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
     3421 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     3481 tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat
     3541 agactgcgtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt
     3601 gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg
     3661 tctttcatgg gtaacagttt cttgaagttg gagaacaaca ttctgagggt aggagtcgaa
     3721 tattaagtaa tcctgactca attagccact gttttgaatc cacatactcc aatactcctg
     3781 aaatagttca ttatggacag cgcagaaaga gctggggaga attgtgaaat tgttatccgc
     3841 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3901 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3961 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4021 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4081 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4141 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4201 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4261 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4321 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4381 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4441 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4501 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4561 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4621 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4681 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4741 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4801 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4861 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4921 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4981 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5041 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5101 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5161 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5221 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5281 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5341 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5401 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5461 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5521 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5581 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5641 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5701 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5761 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5821 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5881 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5941 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6001 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     6061 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     6121 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     6181 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     6241 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     6301 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     6361 ggcgattaag ttgggtaacg ccaggg
//
