LOCUS       sequence-481845-        6389 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  HAT.E.HA Flow Code Plasmid .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6389)
  TITLE     Flow Code Plasmids
REFERENCE   2  (bases 1 to 6389)
  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-12531 HAT.E.HA
FEATURES             Location/Qualifiers
     source          1..6389
                     /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             2952..2978
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             3157..3671
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3833..3849
                     /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(3857..3863,3864..3881,3882..3887))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3902..3923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4040..4057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4211..4799)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4291..4310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4970..5761,5762..5830))
                     /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     5593..5612
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5831..5935)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6003..6021
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6059..6081)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6181..6200
                     /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 agggggggca cctgtgccct atccagatcc cctggagcct aggggaggag
     2941 gcgggggagg gtacccatac gatgtgccag attacgctgg catgctgatg gtgtaagcgg
     3001 ccgcgactct agagtcgacc tgcaggcatg caagcttcag gtagccggct aacgttaaca
     3061 accggtacct ctagaactat agctagcatg cgcaaattta aagcgctgat atcgataaaa
     3121 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     3181 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag
     3241 agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc
     3301 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     3361 ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     3421 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     3481 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc
     3541 gatagactgc gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
     3601 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
     3661 gggtctttca tgggtaacag tttcttgaag ttggagaaca acattctgag ggtaggagtc
     3721 gaatattaag taatcctgac tcaattagcc actgttttga atccacatac tccaatactc
     3781 ctgaaatagt tcattatgga cagcgcagaa agagctgggg agaattgtga aattgttatc
     3841 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3901 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3961 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4021 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4081 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4141 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4201 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4261 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4321 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4381 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4441 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4501 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4561 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4621 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4681 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4741 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4801 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4861 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4921 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4981 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5041 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5101 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5161 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5221 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5281 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5341 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5401 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5461 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5521 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5581 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5641 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5701 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5761 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5821 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5881 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5941 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     6001 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     6061 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     6121 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     6181 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     6241 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     6301 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     6361 caaggcgatt aagttgggta acgccaggg
//
