LOCUS       sequence-373033-        7686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP-tagged DmCCr4 in S2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7686)
  AUTHORS   Haas G, Braun JE, Igreja C, Tritschler F, Nishihara T, Izaurralde E
  TITLE     HPat provides a link between deadenylation and decapping in metazoa.
  JOURNAL   J Cell Biol. 2010 Apr 19. 189(2):289-302.
  PUBMED    20404111
REFERENCE   2  (bases 1 to 7686)
  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: "J Cell 
            Biol. 2010 Apr 19. 189(2):289-302."
COMMENT     Sequence Label: pAc5.1B-EGFP-DmCCr4
FEATURES             Location/Qualifiers
     source          1..7686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..2510
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2530..2532,2533..2535,2536..3246)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2530..2532,2533..2535,2536..3246)
                     /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(2836..2855)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3183..3204
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4937..4978
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4988..5005
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5028..5045)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5078..5212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5102..5121)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5156..5175
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5813..6401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5893..5912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6572..7363,7364..7432))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7195..7214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7433..7537)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7605..7623
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 catgaatggc atcaactctg aatcaaatct ttgcagatgc acctacttct catttccact
       61 gtcacatcat ttttccagat ctcgctgcct gttatgtggc ccacaaacca agacacgttt
      121 tatggccatt aaagctggct gatcgtcgcc aaacaccaaa tacataatga atatgtacac
      181 attcgagaaa gaagcgatca aagaagcgtc ttcgggcgga gtaggagaat gcggaggaga
      241 aggagaacga gctgatctag tatctctcca caatccaatg ccaactgacc aactggccat
      301 attcggagca atttgaagcc aatttccatc gcctggcgat cgctccattc ttggctatat
      361 gtttttcacc gttacccggg gccattttca aagactcgtc ggcaagataa gattgtgtca
      421 ctcgctgtct ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag
      481 tattttgaag aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac
      541 aaaaacatgt tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt
      601 atgtatatac ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac
      661 tgatgatatt ttaagatgat gccagaccaa aaggcttgaa tttctgcgtc ttttgccgaa
      721 cgcagtgcat gtgcaattgt tgttttttgg aatattcaat tttcggactg tccgctttga
      781 tttcagtttc ttggcttatt caaaaagcaa agtaaagcca aaaaagcgag atggcaatac
      841 caaatgcggc aaaacggtag tggaaggaaa ggggtgcggg gcagcggaag gaagggtggg
      901 gcggggcgtg gcggggtctg tggctgggcg cgacgtcacc gacgttggag ccactccttt
      961 gaccatgtgt gcgtgtgtgt attattcgtg tctcgccact cgccggttgt ttttttcttt
     1021 ttatgctgcg ctctctctag cgccatctcg cttacgcatg ctcaacgcac cgcatgttgc
     1081 cgtttccttt tatgcgtcat tttggctcga aataggcaat tatttaaaca aagattagtc
     1141 aacgaaaacg ctaaaataaa taagtctaca atatggttac ttattgccat gtgtgtgcag
     1201 ccaacgatag caacaaaagc aacaacacag gtggctttcc ctctttcact ttttgtttgc
     1261 aagccgcgtg cgagcaagac ggcacgaccg gcaaacgcaa ttacgctgac aaagagcaga
     1321 cgaagttttg gcgaaaaaca tcaaggcgcc tgatacgaat gcatttgcaa taacaattgc
     1381 gatatttaat attgtttatg aagctgtttg acttcaaaac acacaaaaaa aaaaataaaa
     1441 caaattattt gaaagagaat taggaatcgg acgcttatcg ttagggtaac aacaagaaat
     1501 gcttactgag tcacagcctc tggaaaactg ccgcaagcca gagagagaga gaaaaagagg
     1561 gagagcagct tagaccgcat gtgcttgtgt gtgaggcgtc tctctcttcg tctctgttgc
     1621 gcaaacgcat agactgcact gaaaaaatcg attacctatt ttttatgaat gaatatttgc
     1681 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1741 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1801 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1861 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgaatgcc ctactagaag
     1921 atgtgtgttg cacaaaatgt ccctggaata accaatttga agtgcagata gcagtaaacg
     1981 taagctaata tgaatattat ttaactgtaa tgttttaata tcgctggaca ttactaataa
     2041 acccactata aacacatgta catatgtatg ttttggcata caatgagtag ttggggaaaa
     2101 aatgtgtaaa agcaccgtga ccatcacagc ataaagataa ccagctgaag tatcgaatat
     2161 gagtaacccc caaattgaat cacatgccgc aactgatagg acccatggaa gtacactctt
     2221 catggcgata tacaagacac acacaagcac gaacacccag ttgcggagga aattctccgt
     2281 aaatgaaaac ccaatcggcg aacaattcat acccatatat ggtaaaagtt ttgaacgcga
     2341 cttgagagcg gagagcattg cggctgataa ggttttagcg ctaagcgggc tttataaaac
     2401 gggctgcggg accagttttc atatcactac cgtttgagtt cttgtgctgt gtggatactc
     2461 ctcccgacac aaagccgctc catcagccag cagtcgtcta atccagagac cccggatcgg
     2521 ggtaccatca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2581 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     2641 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     2701 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     2761 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2821 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2881 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2941 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     3001 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     3061 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3121 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3181 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3241 tacaagccaa gcttgatgaa aggcaatcat tataaaatgt ctcgtaacaa agacaaatac
     3301 gacagcgcca atcggcggca gcagatcttt ctgtcgcagg aggacatagc tgcgggcaag
     3361 aagacgaact ggagtggcct agagattact ggctgcgtcc gcaatatcag cccgtcgctg
     3421 tgggaatttg agcatctaac cgccctctac ctgaacgaca accagctgct gcgattgccc
     3481 gcagacgttg gcatgctgac cagcctccga actttggacc tgtccagcaa taagctaaga
     3541 agtcttcccg cagagctcgg cgagctcatc cagttgcggg aactgttgct gaacaacaac
     3601 tttctgcgcg tactgcctta cgagatcggc aagctgttcc acctcgtcat actcggcctc
     3661 atgggcaatc cgctgcaaaa ggagttcatg aacatctaca acgaaccgaa cggcacgcag
     3721 aaactgctca cctacatgct ggacaacttg tcatttaccg tgaatccacc gccccagagg
     3781 ccctggctgc cactggccaa gcccaacaaa acgcgaccag cctgcatttt tacggtcatg
     3841 tgctataatg tgctctgcga caagtacgcg acgcgacaaa tgtacggata ctgtccgtcg
     3901 tgggcgctat gctgggagta ccgaaaaaag tcgattatcg acgagatacg gcactatgca
     3961 gcggacatta tcagtctgca ggagatcgaa acggagcaat tctatcactt cttcctgccg
     4021 gaactcaaga acgatggata cgagggaatc ttctcaccga agtcgcgtgc taagactatg
     4081 tccgagctgg agaggaagta cgtcgatggc tgtgcgatat tcttcagggc gtccaagttt
     4141 acgctgatca aggaatcact gatcgagttc aatcagctgg caatggccaa tgccgagggc
     4201 tccgacaaca tgctgaaccg cgtaatgcct aaggataaca tcggtctggc cgcactgctc
     4261 aaggtgaagg agaacgcatg ggagccgatg tccgaggtga cgcagatctc gcagccgctg
     4321 ctcgtctgca cggcgcacat acactgggac cctgagttct gcgacgtcaa gctcatccag
     4381 acgatgatgc ttagcaatga gttgaagacg atcatcgacg aggcgagcca cagtttccga
     4441 cctggtcaca agaacgactc caatgctgtc cagctgctgc tgtgcggtga cttcaactcg
     4501 ctacccgatt caggcgttgt ggagtttctc ggcaagggcc gcgtttccat ggatcatttg
     4561 gacttcaagg acatgggcta caagtcctgc ctgcagcggc tgctctcgaa cgacaccaac
     4621 gagtttacgc actcgttcaa gctagcctcc gcctacaacg aggacatcat gccgcacacc
     4681 aactatacgt tcgattttaa gggcatcatc gactacattt tctacacgaa gacgggcatg
     4741 gtgccgctgg gcctgctggg tcctgtctcc aatgattggc tgcgcgagaa taaggttgtt
     4801 ggatgcccac atccgcatat accctctgat cacttcccac tgctggtcga gctagagctg
     4861 atgcatacgg ctagccaaca ggcgcctcct aacgggctga tcaatcgccg gtaatctaga
     4921 gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     4981 taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     5041 ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat
     5101 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     5161 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     5221 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     5281 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgaccgatg cccttgagag
     5341 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     5401 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt
     5461 tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg
     5521 gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg
     5581 agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt
     5641 tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg
     5701 ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc
     5761 ttcaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5821 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5881 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5941 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6001 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6061 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6121 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6181 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6241 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6301 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6361 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6421 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6481 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6541 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6601 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6661 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6721 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6781 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6841 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg
     6901 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6961 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7021 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7081 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7141 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
     7201 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7261 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7321 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7381 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7441 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7501 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7561 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     7621 ggccctttcg tcttcaagaa ttaattctat attctaaaaa cacaaatgat acttctaaaa
     7681 aaaaat
//
