LOCUS       sequence-473836-        8792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of Hs4E-T.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8792)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8792)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pcDNA3.1-MS2-HA-Hs4E-T_AA
FEATURES             Location/Qualifiers
     source          1..8792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1333..1359
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(4345..4363)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4346..4363)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4383..4400)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4389..4613
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4659..5087
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4746..4765)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4956..4977
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5096..5116)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5101..5430
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5281..5416
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5343..5362
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5497..6291
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5551..5570)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6161..6180
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6465..6586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6502..6521)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6556..6575
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6635..6651)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6635..6651)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6648..6670)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6659..6675
                     /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(6683..6689,6690..6707,6708..6713))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6728..6749
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6866..6883)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7037..7625)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7117..7136)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7796..8587,8588..8656))
                     /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     8419..8438
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8657..8761)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atggcttcta actttactca gttcgttctc gtcgacaatg gcggaactgg cgacgtgact
      961 gtcgccccaa gcaacttcgc taacggggtc gctgaatgga tcagctctaa ctcgcgatca
     1021 caggcttaca aagtaacctg tagcgttcgt cagagctctg cgcagaatcg caaatacacc
     1081 atcaaagtcg aggtgcctaa agtggcaacc cagactgttg gtggtgaaga gcttcctgta
     1141 gccggatgga gatcttactt aaatatggaa ctaaccattc caattttcgc cacgaattcc
     1201 gactgcgagc ttattgttaa ggcaatgcaa ggtctcctaa aagatggaaa cccgattccc
     1261 tcggccatcg cggccaactc cggcatctac ggaggtggag gtggatctgg tggatctcct
     1321 ccacatatgg cttacccata cgatgttcca gattacgctc ctccaaagct tggatccatg
     1381 gataggagaa gtatgggtga aacagaaagt ggagatgctt tccttgacct gaagaagcct
     1441 cctgcctcca aatgccccca tcgctataca aaagaagaac tcttggatat aaaagaactc
     1501 ccccattcca aacagaggcc ttcatgcctt tctgaaaaat atgacagtga tggtgtctgg
     1561 gaccctgaga agtggcatgc ctctctctac ccagcttcag ggcggagctc accagtggaa
     1621 agtctgaaga aagagttgga tacagaccgg ccttccctgg tgcgcaggat agtagatcca
     1681 cgagagcgtg tgaaagaaga tgacttagat gttgttctca gccctcagag acggagcttt
     1741 ggagggggct gccacgtgac agccgctgtt agctcccggc gctcaggaag tccattagag
     1801 aaagatagtg atgggcttcg tctgcttggt ggacgtagga ttggcagtgg gaggataatc
     1861 tctgcccgga cctttgagaa ggatcaccgt cttagcgata aggacctgcg ggacttgaga
     1921 gacagagacc gagagaggga cttcaaggac aagcgtttca ggagagagtt tggagatagt
     1981 aagcgtgtct ttggtgagcg tagaagaaat gattcttaca cagaagaaga accagagtgg
     2041 ttctctgctg gacccacaag tcagtctgaa accatcgaac tgactggctt tgatgataag
     2101 atactagaag aagatcacaa agggagaaaa agaacaaggc gacggacagc ctctgtgaag
     2161 gaaggtatag tagagtgcaa tggaggagtg gccgaagagg atgaagtgga ggtcatcctt
     2221 gcacaggagc ctgcggctga tcaggaagtg ccaagggatg ctgtcttgcc tgagcagtcc
     2281 ccaggagact ttgactttaa tgagttcttt aaccttgata aggtgccatg cttggcttcg
     2341 atgatagaag atgttttggg agaagggtca gtctctgcca gtcggttcag taggtggttc
     2401 tctaacccga gcagatcagg aagccgatcc agcagtcttg ggtcaacacc acatgaagag
     2461 ctagagagac ttgcaggtct ggagcaagcc atcctctctc ctggacagaa ctcggggaat
     2521 tactttgctc ctataccatt ggaagaccat gctgaaaata aagtggatat tttagaaatg
     2581 ctacagaaag ccaaagtgga tttgaaacct cttctttcca gcctttctgc aaataaagaa
     2641 aaacttaaag aaagctcaca ttcaggggtt gtgctttcag tggaggaggt agaagcaggt
     2701 ctgaagggct tgaaggttga ccagcaagtg aagaattcaa ctcccttcat ggcagaacac
     2761 ctagaagaga ccttgagtgc cgtaaccaac aatcgacaac tgaagaaaga cggagacatg
     2821 actgcgttca acaagctagt gagcacaatg aaggcaagtg ggactttgcc ttctcagccc
     2881 aaagtcagcc gaaaccttga aagccatttg atgtcccctg ctgagattcc aggccagcct
     2941 gtccctaaga acatcctgca ggaacttctg ggtcaaccag ttcagagacc tgcttcttcc
     3001 aatcttctga gtggccttat ggggagcttg gagcctacaa catctttact gggccaaaga
     3061 gcaccctctc ctcccttgtc acaggtgttt caaactcgag cagcctcagc tgactacctt
     3121 cgcccaagaa taccatcacc aattggtttc acaccaggac cacagcagct actcggagat
     3181 ccattccaag gcatgcgcaa acccatgagc cccatcacag cccagatgag ccagctggag
     3241 ttgcaacagg cagctttaga agggctggcc ttgccacatg accttgctgt acaggcagca
     3301 aacttctacc agcctggttt tggcaaacca caggtggaca gaaccagaga tggattcaga
     3361 aacaggcaac agcgagtgac caagtcacca gcacccgtgc atcgagggaa ttcctcttcc
     3421 cctgcccctg ctgcctccat cacaagcatg ctttctcctt cctttacccc tacctcagtg
     3481 attcgtaaga tgtacgagag caaagagaaa agcaaggagg agccagcatc tggaaaagca
     3541 gctcttggtg acagtaaaga ggatactcag aaggccagtg aagaaaacct cctgtcatcc
     3601 agctctgtac ccagtgccga tcgagactct tctcccacta caaattccaa actgtcagca
     3661 ttacagaggt cttcgtgttc caccccactg tcccaggcca accgttacac caaagaacaa
     3721 gattatcgac ctaaagcaac tgggagaaaa acacccacct tggcatcccc agttcctaca
     3781 acaccttttc tccgccctgt ccaccaagtt ccccttgtcc cccatgtccc tatggttagg
     3841 cctgctcacc agcttcaccc agggttggta cagaggatgc tggcccaggg agtacatcca
     3901 cagcatcttc caagtttgct ccaaactggt gtgcttcctc ctgggatgga cttgagtcat
     3961 ttacagggaa tatctggccc catcctgggt cagccctttt accctttacc tgctgctagt
     4021 caccctctct taaaccctcg tcctggaaca cctctgcatc tggcaatggt gcaacagcag
     4081 ctacagcgct cagttctgca tcctccaggc tctggttccc atgcagcagc tgtcagcgtt
     4141 cagacaaccc ctcagaacgt gcccagccgg tcaggcctgc cccacatgca ctcccagctg
     4201 gagcatcgcc ccagccagag gagcagctcc cctgtgggcc ttgccaaatg gtttggctca
     4261 gatgtgctac agcaacccct gccctccatg cccgccaaag ttatcagtgt agatgaattg
     4321 gaataccgac agtgagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     4381 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4441 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4501 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4561 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     4621 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4681 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4741 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4801 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4861 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4921 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4981 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     5041 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     5101 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5161 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5221 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5281 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5341 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     5401 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     5461 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5521 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5581 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5641 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5701 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5761 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5821 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5881 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5941 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     6001 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     6061 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     6121 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     6181 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6241 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6301 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     6361 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6421 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     6481 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6541 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6601 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6661 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6721 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6781 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6841 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6901 tgcggcgagc ggtatcagcc cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6961 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7021 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7081 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7141 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7201 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7261 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7321 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7381 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7441 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7501 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7561 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7621 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7681 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7741 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7801 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7861 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7921 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7981 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8041 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8101 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8161 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8221 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8281 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8341 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8401 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8461 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8521 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8581 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8641 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8701 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8761 gcacatttcc ccgaaaagtg ccacctgacg tc
//
