LOCUS       sequence-481840-        6395 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  HAT.OLLAS.NWS Flow Code Plasmid .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6395)
  TITLE     Flow Code Plasmids
REFERENCE   2  (bases 1 to 6395)
  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-12495 HAT.OLLAS.NWS
FEATURES             Location/Qualifiers
     source          1..6395
                     /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..2936
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2961..2984
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     LTR             3163..3677
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3839..3855
                     /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(3863..3869,3870..3887,3888..3893))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3908..3929
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4046..4063)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4217..4805)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4297..4316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4976..5767,5768..5836))
                     /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     5599..5618
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5837..5941)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6009..6027
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6065..6087)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6187..6206
                     /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 agggagcggc ttcgccaacg agctgggccc caggctgatg ggcaagtgcg
     2941 gaggaggcgg gggagggaac tggtcccacc cccaattcga gaaaggcatg ctgatggtgt
     3001 aagcggccgc gactctagag tcgacctgca ggcatgcaag cttcaggtag ccggctaacg
     3061 ttaacaaccg gtacctctag aactatagct agcatgcgca aatttaaagc gctgatatcg
     3121 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     3181 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat acataactga
     3241 gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag
     3301 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     3361 gcggtcccgc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     3421 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     3481 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag
     3541 tcctccgata gactgcgtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     3601 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     3661 cagcgggggt ctttcatggg taacagtttc ttgaagttgg agaacaacat tctgagggta
     3721 ggagtcgaat attaagtaat cctgactcaa ttagccactg ttttgaatcc acatactcca
     3781 atactcctga aatagttcat tatggacagc gcagaaagag ctggggagaa ttgtgaaatt
     3841 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     3901 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     3961 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4021 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4081 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4141 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4201 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4261 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4321 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4381 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4441 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4501 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4561 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4621 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4681 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4741 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4801 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4861 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4921 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4981 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5041 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5101 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5161 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5221 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5281 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5341 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5401 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5461 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5521 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5581 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5641 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5701 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5761 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5821 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5881 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5941 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     6001 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     6061 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     6121 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     6181 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     6241 acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact
     6301 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     6361 gtgctgcaag gcgattaagt tgggtaacgc caggg
//
