LOCUS       sequence-386576-        6679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPABPC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6679)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6679)
  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-HsPABPC1_I
FEATURES             Location/Qualifiers
     source          1..6679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     553..572
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        553..571
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(605..607,608..610,611..1321)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(605..607,608..610,611..1321)
                     /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(650..671)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(911..930)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1258..1279
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3407..3528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3444..3463)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3498..3517
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3535..3990)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3672..3693)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3884..3903
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4017..4121
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4123..4480
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4123..4314
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4146..4166)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4331..4466
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4515..5309
                     /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             4515..5309
                     /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(4569..4588)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5179..5198
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5497..5516)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5541..5588
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5917..6505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6406..6425
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      541 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      601 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      661 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      721 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      781 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      841 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      901 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
      961 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1021 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1081 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1141 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1201 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1261 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1321 gtccggactc agatctcgag ctcaagcttc gaattctgca gtcgacatga accccagtgc
     1381 ccccagctac cccatggcct cgctctacgt gggggacctc caccccgacg tgaccgaggc
     1441 gatgctctac gagaagttca gcccggccgg gcccatcctc tccatccggg tctgcaggga
     1501 catgatcacc cgccgctcct tgggctacgc gtatgtgaac ttccagcagc cggcggacgc
     1561 ggagcgtgct ttggacacca tgaattttga tgttataaag ggcaagccag tacgcatcat
     1621 gtggtctcag cgtgatccat cacttcgcaa aagtggagta ggcaacatat tcattaaaaa
     1681 tctggacaaa tccattgata ataaagcact gtatgataca ttttctgctt ttggtaacat
     1741 cctttcatgt aaggtggttt gtgatgaaaa tggttccaag ggctatggat ttgtacactt
     1801 tgagacgcag gaagcagctg aaagagctat tgaaaaaatg aatggaatgc tcctaaatga
     1861 tcgcaaagta tttgttggac gatttaagtc tcgtaaagaa cgagaagctg aacttggagc
     1921 tagggcaaaa gaattcacca atgtttacat caagaatttt ggagaagaca tggatgatga
     1981 gcgccttaag gatctctttg gcaagtttgg gcctgcctta agtgtgaaag taatgactga
     2041 tgaaagtgga aaatccaaag gatttggatt tgtaagcttt gaaaggcatg aagatgcaca
     2101 gaaagctgtg gatgagatga acggaaagga gctcaatgga aaacaaattt atgttggtcg
     2161 agctcagaaa aaggtggaac ggcagacgga acttaagcgc aaatttgaac agatgaaaca
     2221 agataggatc accagatacc agggtgttaa tctttatgtg aaaaatcttg atgatggtat
     2281 tgatgatgaa cgtctccgga aagagttttc tccatttggt acaatcacta gtgcaaaggt
     2341 tatgatggag ggtggtcgca gcaaagggtt tggttttgta tgtttctcct ccccagaaga
     2401 agccactaaa gcagttacag aaatgaacgg tagaattgtg gccacaaagc cattgtatgt
     2461 agctttagct cagcgcaaag aagagcgcca ggctcacctc actaaccagt atatgcagag
     2521 aatggcaagt gtacgagctg ttcccaaccc tgtaatcaac ccctaccagc cagcacctcc
     2581 ttcaggttac ttcatggcag ctatcccaca gactcagaac cgtgctgcat actatcctcc
     2641 tagccaaatt gctcaactaa gaccaagtcc tcgctggact gctcagggtg ccagacctca
     2701 tccattccaa aatatgcccg gtgctatccg cccagctgct cctagaccac catttagtac
     2761 tatgagacca gcttcttcac aggttccacg agtcatgtca acacagcgtg ttgctaacac
     2821 atcaacacag acaatgggtc cacgtcctgc agctgcagcc gctgcagcta ctcctgctgt
     2881 ccgcaccgtt ccacagtata aatatgctgc aggagttcgc aatcctcagc aacatcttaa
     2941 tgcacagcca caagttacaa tgcaacagcc tgctgttcat gtacaaggtc aggaaccttt
     3001 gactgcttcc atgttggcat ctgcccctcc tcaagagcaa aagcaaatgt tgggtgaacg
     3061 gctgtttcct cttattcaag ccatgcaccc tactcttgct ggtaaaatca ctggcatgtt
     3121 gttggagatt gataattcag aacttcttca tatgctcgag tctccagagt cactccgttc
     3181 taaggttgat gaagctgtag ctgtactaca agcccaccaa gctaaagagg ctgcccagaa
     3241 agcagttaac agtgccaccg gtgttccaac tgtttaagga tccaccggat ctagataact
     3301 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3361 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3421 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3481 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg
     3541 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3601 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3661 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3721 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3781 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3841 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3901 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3961 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     4021 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4081 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     4141 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4201 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4261 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4321 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4381 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4441 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     4501 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4561 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4621 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4681 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4741 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4801 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4861 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4921 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4981 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5041 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5101 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5161 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5221 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5281 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5341 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5401 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5461 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     5521 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     5581 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     5641 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     5701 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     5761 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5821 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5881 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5941 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6001 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6061 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6121 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6181 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6241 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6301 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6361 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6421 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6481 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6541 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6601 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata
     6661 gcccatatat ggagttccg
//
