LOCUS       sequence-289776-        6838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmaug2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6838)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6838)
  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: pT7-EGFP-C1-HsSmaug2_Q
FEATURES             Location/Qualifiers
     source          1..6838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(710..731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(971..990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1318..1339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3626..3747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3663..3682)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3717..3736
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3754..4209)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3891..3912)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4103..4122
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4236..4340
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4342..4699
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4342..4533
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4365..4385)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4550..4685
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4734..5528
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4734..5528
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4788..4807)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5398..5417
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5716..5735)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5760..5807
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6136..6724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6625..6644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1381 gtccggactc agatctcgag ctcaagcttc catgatgttc cgagaccagg tgggcatcct
     1441 cgctggctgg ttcaaaggct ggaatgagtg tgagcagaca gtggccctcc tgtcacttct
     1501 gaaacgggtc acccgtaccc aggcccgctt cctgcagctc tgcctggagc actcactggc
     1561 ggactgcaat gacatccacc tgctggagtc ggaggccaac agtgctgcca tcgtcagcca
     1621 gtggcagcag gagtccaaag agaaggtggt gtccctcctg ctgtcccacc ttcccctgct
     1681 tcagccaggc aacacagagg ccaagtcgga gtacatgagg ctactgcaga aagtgctggc
     1741 ctactcaatc gagagcaatg ctttcatcga ggagagtcgc cagctgcttt cctatgccct
     1801 catccaccca gccaccacac tggaggaccg caacgcactg gccctctggc tgagccacct
     1861 ggaagagcgg ttggctagtg gcttccgctc ccggccagag ccctcctacc attcacgtca
     1921 aggctcagat gagtgggggg gccctgcaga gctaggccct ggggaggcag ggccaggctg
     1981 gcaggacaag ccaccccggg aaaatggaca cgtgcccttc cacccatcca gctcagtgcc
     2041 gccagccatc aacagtattg ggagcaatgc aaacacaggt ctcccctgcc aaatccaccc
     2101 tagcccactg aagcgctcca tgtcactcat ccctacaagc ccccaggtcc ctggtgagtg
     2161 gccgagtcca gaggagcttg gggcccgggc tgcttttacc acgcccgatc acgcacctct
     2221 ctcgccccag agcagcgtgg cctcctctgg cagtgagcag acagaggagc agggctccag
     2281 ccggaacacc ttccaggagg atggcagtgg catgaaagat gtgccctcat ggctcaagag
     2341 cctccgtttg cacaagtatg cagccctctt ctcacagatg agctacgagg agatgatgac
     2401 actgactgag cagcacctgg agtctcagaa cgtcaccaaa ggtgcccgcc acaagatagc
     2461 cctgagcatc cagaagctgc gtgagagaca gagcgtcctc aagtccctag agaaggatgt
     2521 gctggaaggc gggaacctac gaaacgctct gcaggagctg cagcagatca tcatcactcc
     2581 catcaaggcc tacagtgtcc tccaggccac cgtggctgcc gccaccacca cccctactgc
     2641 caaggatggg gccccggggg aaccaccgct gccaggtgct gagcctcccc tagcccaccc
     2701 cggcacagac aaaggcaccg aggccaagga ccctccagct gtggagaact acccacctcc
     2761 accagctcca gctcccactg atggcagtga gcctgccccg gctcccgtcg ccgacggaga
     2821 catccccagc cagtttacac gggtgatggg caaagtgtgc acccaactgc tggtgtcccg
     2881 accagacgag gagaacatca ccagttacct ccagctcatc gaaaagtgcc tgactcatga
     2941 ggctttcacg gagacgcaga agaaacggct gctatcctgg aaacagcaag tgctgaagct
     3001 cctccggaca ttcccgcgca aagccgcact agagatgcag aactaccggc agcagaaagg
     3061 ctgggcattc ggctccaact cgctccccat agctggctct gtggggatgg gagtggcccg
     3121 gcgtacccag cggcagttcc caatgcctcc ccgggccctc ccacccggcc ggatgggcct
     3181 cctgagcccc tcgggcattg ggggtgtctc ccctcgacat gccctcacca gccccagcct
     3241 tggaggccag ggccgacaga acctgtggtt tgccaaccct ggaggcagca acagcatgcc
     3301 cagtcagagc cgcagctctg tgcagcgcac ccactcgctc ccggtccact cgtcacccca
     3361 ggccattctc atgttccctc cagactgccc ggtccctggg cctgacctgg agatcaatcc
     3421 cactctggag tctctgtgtc tgagcatgac agaacacgcc ttgggtgatg ggacagacaa
     3481 aacctccacc atctgaggat ccaccggatc tagataactg atcataatca gccataccac
     3541 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     3601 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     3661 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3721 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa
     3781 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3841 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3901 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3961 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4021 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4081 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4141 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4201 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4261 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4321 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     4381 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4441 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4501 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4561 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4621 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4681 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     4741 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4801 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4861 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     4921 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4981 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5041 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5101 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5161 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5221 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     5281 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5341 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5401 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5461 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5521 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5581 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5641 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5701 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     5761 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     5821 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     5881 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     5941 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     6001 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6061 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6121 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6181 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6241 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     6301 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6361 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6421 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6481 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6541 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6601 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6661 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6721 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6781 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
