LOCUS       sequence-299557-        6841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-TTP in mammalian cells, can be used to make inducible 
            cell line.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6841)
  AUTHORS   Tollenaere MAX, Tiedje C, Rasmussen S, Nielsen JC, Vind AC, Blasius 
            M, Batth TS, Mailand N, Olsen JV, Gaestel M, Bekker-Jensen S
  TITLE     GIGYF1/2-Driven Cooperation between ZNF598 and TTP in 
            Posttranscriptional Regulation of Inflammatory Signaling.
  JOURNAL   Cell Rep. 2019 Mar 26;26(13):3511-3521.e4. doi: 
            10.1016/j.celrep.2019.03.006.
  PUBMED    30917308
REFERENCE   2  (bases 1 to 6841)
  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"; doi: 
            "10.1016/j.celrep.2019.03"; journalName: "Cell Rep"; date: 
            "2019-03-26- 26"; volume: "26"; issue: "13"; pages: "3511-3521.e4"
COMMENT     Sequence Label: pcDNA4/TO/GFP-TTP
FEATURES             Location/Qualifiers
     source          1..6841
                     /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"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(984..986,987..989,990..1697)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(984..986,987..989,990..1697)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(1290..1309)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1637..1658
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1716..1740
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2731..2755)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2849..2866)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2855..3079
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3125..3553
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3212..3231)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3422..3443
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3562..3582)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3567..3896
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3747..3882
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3809..3828
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3944..3991
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4010..4384
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4514..4635
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4551..4570)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4605..4624
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4684..4700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4684..4700)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4697..4719)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4708..4724
                     /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(4732..4738,4739..4756,4757..4762))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4777..4798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4915..4932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5086..5674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5166..5185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5845..6636,6637..6705))
                     /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     6468..6487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6706..6810)
                     /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 ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttatggtga gcaagggcga ggagctgttc accggggtgg
     1021 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1081 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1141 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1201 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1261 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1321 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1381 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1441 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1501 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1561 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     1621 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1681 gcatggacga gctgtacggt accgagctcg gatctacaag tttgtacaaa aaagcaggct
     1741 ccaccatgga tctgactgcc atctacgaga gcctcctgtc gctgagccct gacgtgcccg
     1801 tgccatccga ccatggaggg actgagtcca gcccaggctg gggctcctcg ggaccctgga
     1861 gcctgagccc ctccgactcc agcccgtctg gggtcacctc ccgcctgcct ggccgctcca
     1921 ccagcctagt ggagggccgc agctgtggct gggtgccccc accccctggc ttcgcaccgc
     1981 tggctccccg cctgggccct gagctgtcac cctcacccac ttcgcccact gcaacctcca
     2041 ccaccccctc gcgctacaag actgagctat gtcggacctt ctcagagagt gggcgctgcc
     2101 gctacggggc caagtgccag tttgcccatg gcctgggcga gctgcgccag gccaatcgcc
     2161 accccaaata caagacggaa ctctgtcaca agttctacct ccagggccgc tgcccctacg
     2221 gctctcgctg ccacttcatc cacaacccta gcgaagacct ggcggccccg ggccaccctc
     2281 ctgtgcttcg ccagagcatc agcttctccg gcctgccctc tggccgccgg acctcaccac
     2341 caccaccagg cctggccggc ccttccctgt cctccagctc cttctcgccc tccagctccc
     2401 caccaccacc tggggacctt ccactgtcac cctctgcctt ctctgctgcc cctggcaccc
     2461 ccctggctcg aagagacccc accccagtct gttgcccctc ctgccgaagg gccactccta
     2521 tcagcgtctg ggggcccttg ggtggcctgg ttcggacccc ctctgtacag tccctgggat
     2581 ccgaccctga tgaatatgcc agcagcggca gcagcctggg gggctctgac tctcccgtct
     2641 tcgaggcggg agtttttgca ccaccccagc ccgtggcagc cccccggcga ctccccatct
     2701 tcaatcgcat ctctgtttct gagtggatcc acccagcttt cttgtacaaa gtggtagatc
     2761 cactagtcca gtgtggtgga attctgcaga tatccagcac agtggcggcc gctcgagtct
     2821 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2881 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2941 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3001 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3061 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3121 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3181 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3241 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3301 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3361 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3421 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3481 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3541 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3601 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3661 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3721 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3781 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3841 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3901 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc
     3961 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt
     4021 gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac
     4081 cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga
     4141 cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc
     4201 ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac
     4261 gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg
     4321 ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga
     4381 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4441 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4501 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4561 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4621 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4681 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4741 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4801 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4861 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4921 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4981 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5041 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5101 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5161 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5221 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5281 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5341 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5401 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5461 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5521 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5761 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5821 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5881 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5941 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6001 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6061 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6121 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6181 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6241 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6301 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6361 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6421 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6481 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6541 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6601 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6661 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6721 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6841 c
//
