LOCUS       sequence-254828-        8892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing codon-optimized GFP(1-10) fragment in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8892)
  AUTHORS   Salo VT, Li S, Vihinen H, Holtta-Vuori M, Szkalisity A, Horvath P, 
            Belevich I, Peranen J, Thiele C, Somerharju P, Zhao H, Santinho A, 
            Thiam AR, Jokitalo E, Ikonen E
  TITLE     Seipin Facilitates Triglyceride Flow to Lipid Droplet and 
            Counteracts Droplet Ripening via Endoplasmic Reticulum Contact.
  JOURNAL   Dev Cell. 2019 Jun 6. pii: S1534-5807(19)30388-0. doi: 
            10.1016/j.devcel.2019.05.016.
  PUBMED    31178403
REFERENCE   2  (bases 1 to 8892)
  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"; journalName: "Dev Cell. 
            2019 Jun 6. pii: S1534-5807(19)30388-0. doi: 
            10.1016/j.devcel.2019.05.016."
COMMENT     Sequence Label: pSH-EFIRES-P-GFP(1-10)opti
FEATURES             Location/Qualifiers
     source          1..8892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(135..152)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(306..894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(386..405)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1065..1856,1857..1925))
                     /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     1688..1707
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1926..2030)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2057..2512
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2144..2163)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2354..2375
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    3029..3832
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     polyA_signal    3858..3906
                     /note="synthetic polyadenylation signal"
     misc_feature    3920..4011
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3960..3979
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        4023..5185
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4254..5176
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5133..5153
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     intron          5224..5356
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5339..5358
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        5401..5419
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    6133..6699
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    6149..6701
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6293..6310)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6520..6539
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6701..7303
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7200..7220
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7537..7618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7574..7593)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    7646..8482
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
ORIGIN
        1 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta
       61 tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag
      121 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
      181 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
      241 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      301 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      361 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      421 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      481 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      541 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      601 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
      661 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
      721 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
      781 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
      841 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
      901 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
      961 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     1021 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     1081 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     1141 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     1201 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     1261 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     1321 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     1381 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     1441 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     1501 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     1561 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     1621 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     1681 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     1741 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     1801 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     1861 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     1921 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     1981 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     2041 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2101 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2161 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2221 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2281 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2341 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2401 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2461 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttgccattcg
     2521 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     2581 cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact tggagcggcc
     2641 gtcgacctac tctcttccgc attggagtcg ctttaactgg ccctggcttt ggcagcctgt
     2701 gctgacccat gcagtcctcc ttaccatccc tccctcgact tcccctcttc cgatgttgag
     2761 cccctccagc cggtcctgga ctttgtctcc ttccctgccc tgccctctcc tgaacctgag
     2821 ccagctccca tagctcagtc tggtctatct gcctggccct ggccattgtc actttgcgct
     2881 gccctcctct cgcccccgag tgcccttgct gtgccgccgg aactctgccc tctaacgctg
     2941 ccgtctctct cctgagtccg gaccactttg agctctactg gcttctgcgc cgcctctggc
     3001 ccactgtttc cccttcccag gcaggtcctg ctttctctga cctgcattct ctcccctggg
     3061 cctgtgccgc tttctgtctg cagcttgtgg cctgggtcac ctctacggct ggcccagatc
     3121 cttccctgcc gcctccttca ggttccgtct tcctccactc cctcttcccc ttgctctctg
     3181 ctgtgttgct gcccaaggat gctctttccg gagcacttcc ttctcggcgc tgcaccacgt
     3241 gatgtcctct gagcggatcc tccccgtgtc tgggtcctct ccgggcatct ctcctccctc
     3301 acccaacccc atgccgtctt cactcgctgg gttccctttt ccttctcctt ctggggcctg
     3361 tgccatctct cgtttcttag gatggccttc tccgacggat gtctcccttg cgtcccgcct
     3421 ccccttcttg taggcctgca tcatcaccgt ttttctggac aaccccaaag taccccgtct
     3481 ccctggcttt agccacctct ccatcctctt gctttctttg cctggacacc ccgttctcct
     3541 gtggattcgg gtcacctctc actcctttca tttgggcagc tcccctaccc cccttacctc
     3601 tctagtctgt gctagctctt ccagccccct gtcatggcat cttccagggg tccgagagct
     3661 cagctagtct tcttcctcca acccgggccc ctatgtccac ttcaggacag catgtttgct
     3721 gcctccaggg atcctgtgtc cccgagctgg gaccacctta tattcccagg gccggttaat
     3781 gtggctctgg ttctgggtac ttttatctgt cccctccacc ccacagtggg gcgcatgcgt
     3841 ggaattatca cctcgagaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     3901 tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac
     3961 tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc tactagtcgt
     4021 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     4081 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     4141 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     4201 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     4261 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     4321 ttacttccac ctggctccag tacgtgattc ttgatcccga gctggagcca ggggcgggcc
     4381 ttgcgcttta ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
     4441 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
     4501 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
     4561 aaatgcgggc caggatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
     4621 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
     4681 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
     4741 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
     4801 aagatggccg cttcccggcc ctgctccagg gggctcaaaa tggaggacgc ggcgctcggg
     4861 agagcgggcg ggtgagtcac ccacacaaag gaaaggggcc tttccgtcct cagccgtcgc
     4921 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctggagctt
     4981 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
     5041 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
     5101 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
     5161 ttttcttcca tttcaggtgt cgtgaacacg ggctgcagaa gttggtcgtg aggcactggg
     5221 caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg
     5281 agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt
     5341 gcctttctct ccacaggtgt ccactcccag ttcaattaca gctcttaagg ctagagtact
     5401 taatacgact cactataggc tagcgctacc ggactcagat ctaccatggg gtcaaaaggc
     5461 gaagagctgt ttactggggt cgtccctatt ctggtggaac tggatggcga tgtcaatggg
     5521 cataagttta gcgtgagggg agagggagag ggcgacgcaa ccatcggcaa gctgacactg
     5581 aagttcatct gcaccacagg caagctgccc gtgccttggc caaccctggt gaccacactg
     5641 acatacggcg tgcagtgttt tagccggtat ccagaccaca tgaagagaca cgatttcttt
     5701 aagagcgcca tgcccgaggg ctacgtgcag gagaggacaa tctccttcaa ggacgatggc
     5761 aagtataaga cccgcgccgt ggtgaagttt gagggcgaca cactggtgaa ccggatcgag
     5821 ctgaagggca ccgacttcaa ggaggatggc aatatcctgg gccacaagct ggagtacaac
     5881 tttaattctc acaacgtgta tatcacagcc gataagcaga agaacggcat caaggccaat
     5941 ttcaccgtgc ggcacaatgt ggaggacggc agcgtgcagc tggccgatca ctaccagcag
     6001 aacaccccta tcggcgacgg acccgtgctg ctgcctgata atcactatct gtccacccag
     6061 actgtgctgt caaaggaccc aaatgagaaa gggaccggag ggggaggaag ctagcggccg
     6121 ctctagccca attccgcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     6181 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     6241 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     6301 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     6361 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     6421 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     6481 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     6541 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     6601 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     6661 cacggggacg tggttttcct ttgaaaaaca cgatgataat atgggcaccg agtacaagcc
     6721 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     6781 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     6841 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     6901 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     6961 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     7021 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     7081 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     7141 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     7201 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     7261 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc
     7321 gcagcgcccg accgaaagga gcgcacgacc ccatggctcc gaccgaagcc acccggggcg
     7381 gccccgccga ccccgcaccc gcccccgagg cccaccgact ctagctagag gatcataatc
     7441 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     7501 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     7561 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     7621 tctagttgtg gtttgtccat gcatcactag ggacaggatt ggtgacagaa aagccccatc
     7681 cttaggcctc ctccttccta gtctcctgat attgggtcta acccccacct cctgttaggc
     7741 agattcctta tctggtgaca cacccccatt tcctggagcc atctctctcc ttgccagaac
     7801 ctctaaggtt tgcttacgat ggagccagag aggatcctgg gagggagagc ttggcagggg
     7861 gtgggaggga agggggggat gcgtgacctg cccggttctc agtggccacc ctgcgctacc
     7921 ctctcccaga acctgagctg ctctgacgcg gctgtctggt gcgtttcact gatcctggtg
     7981 ctgcagcttc cttacacttc ccaagaggag aagcagtttg gaaaaacaaa atcagaataa
     8041 gttggtcctg agttctaact ttggctcttc acctttctag tccccaattt atattgttcc
     8101 tccgtgcgtc agttttacct gtgagataag gccagtagcc agccccgtcc tggcagggct
     8161 gtggtgagga ggggggtgtc cgtgtggaaa actccctttg tgagaatggt gcgtcctagg
     8221 tgttcaccag gtcgtggccg cctctactcc ctttctcttt ctccatcctt ctttccttaa
     8281 agagtcccca gtgctatctg ggacatattc ctccgcccag agcagggtcc cgcttcccta
     8341 aggccctgct ctgggcttct gggtttgagt ccttggcaag cccaggagag gcgctcaggc
     8401 ttccctgtcc cccttcctcg tccaccatct catgcccctg gctctcctgc cccttcccta
     8461 caggggttcc tggctctgct cttcagactg agccccgttc ccctgcatcc ccgttcccct
     8521 gcatccccct tcccctgcat cccccagagg ccccaggcca cctacttggc ctggacccca
     8581 cgagaggcca ccccagccct gtctaccagg ctgccttttg ggtggattct cctccaactg
     8641 tggggtgact gcttggcaaa ctcactcttc ggggtatccc aggaggcctg gagcattggg
     8701 gtgggctggg gttcagagag gagggattcc cttctcaggt tacgtggcca agaagcaggg
     8761 gagctgggtt tgggtcaggt ctgggtgtgg ggtgaccagc ttatgctgtt tgcccaggac
     8821 agcctagttt tagcgctgaa accctcagtc ctaggaaaac agggatggtt ggtcactgtc
     8881 tctgggctcg ac
//
