LOCUS       sequence-268595-        6501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the P4 coiled coil fused to the ABI abscisic 
            acid-inducible domain in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6501)
  AUTHORS   Lebar T, Lainscek D, Merljak E, Aupic J, Jerala R
  TITLE     A tunable orthogonal coiled-coil interaction toolbox for engineering
            mammalian cells.
  JOURNAL   Nat Chem Biol. 2020 Jan 6. pii: 10.1038/s41589-019-0443-y. doi: 
            10.1038/s41589-019-0443-y.
  PUBMED    31907374
REFERENCE   2  (bases 1 to 6501)
  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: "Nat Chem 
            Biol. 2020 Jan 6. pii: 10.1038/s41589-019-0443-y. doi: 
            10.1038/s41589-019-0443-y."
COMMENT     Sequence Label: pcDNA3-Myc:P4:gs:nls:ABI
FEATURES             Location/Qualifiers
     source          1..6501
                     /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"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      944..953
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             953..982
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1115..1135
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(2054..2072)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2055..2072)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2092..2109)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2098..2322
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2368..2796
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2455..2474)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2665..2686
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2805..2825)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2810..3139
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2990..3125
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3052..3071
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3206..4000
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3260..3279)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3870..3889
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4174..4295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4211..4230)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4265..4284
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4344..4360)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4344..4360)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4357..4379)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4368..4384
                     /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(4392..4398,4399..4416,4417..4422))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4437..4458
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4575..4592)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4746..5334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4826..4845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5505..6296,6297..6365))
                     /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     6128..6147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6366..6470)
                     /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 ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcgccacca tggagcagaa
      961 gctgatcagc gaggaggacc tgagcccgga agataaaatt gctcagctga aacaaaaaat
     1021 ccaagcgctg aaacaggaaa accagcagct ggaagaggaa aacgccgcac tggaatatgg
     1081 tggcggctct ggcggcggct ccggaggctc tgatccaaaa aagaagagaa aggtaacgcg
     1141 tgtgcctttg tatggtttta cttcgatttg tggaagaaga cctgagatgg aagctgctgt
     1201 ttcgactata ccaagattcc ttcaatcttc ctctggttcg atgttagatg gtcggtttga
     1261 tcctcaatcc gccgctcatt tcttcggtgt ttacgacggc catggcggtt ctcaggtagc
     1321 gaactattgt agagagagga tgcatttggc tttggcggag gagatagcta aggagaaacc
     1381 gatgctctgc gatggtgata cgtggctgga gaagtggaag aaagctcttt tcaactcgtt
     1441 cctgagagtt gactcggaga ttgagtcagt tgcgccggag acggttgggt caacgtcggt
     1501 ggttgccgtt gttttcccgt ctcacatctt cgtcgctaac tgcggtgact ctagagccgt
     1561 tctttgccgc ggcaaaactg cacttccatt atccgttgac cataaaccgg atagagaaga
     1621 tgaagctgcg aggattgaag ccgcaggagg gaaagtgatt cagtggaatg gagctcgtgt
     1681 tttcggtgtt ctcgccatgt cgagatccat tggcgataga tacttgaaac catccatcat
     1741 tcctgatccg gaagtgacgg ctgtgaagag agtaaaagaa gatgattgtc tgattttggc
     1801 gagtgacggg gtttgggatg taatgacgga tgaagaagcg tgtgagatgg caaggaagcg
     1861 gattctcttg tggcacaaga aaaacgcggt ggctggggat gcatcgttgc tcgcggatga
     1921 gcggagaaag gaagggaaag atcctgcggc gatgtccgcg gctgagtatt tgtcaaagct
     1981 ggcgatacag agaggaagca aagacaacat aagtgtggtg gtggttgatt tgaagtaatc
     2041 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     2101 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2161 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2221 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2281 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2341 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2401 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2461 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2521 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2581 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccttttga
     2641 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2701 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2761 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     2821 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2881 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2941 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3001 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3061 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3121 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     3181 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3241 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3301 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3361 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3421 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3481 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3541 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3601 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3661 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3721 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3781 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     3841 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     3901 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     3961 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4021 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4081 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4141 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4201 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4261 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4321 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4381 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4441 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4501 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4561 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4621 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4681 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4741 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4801 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4861 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4921 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4981 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5041 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5101 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5161 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5221 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5281 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5341 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5401 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5461 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5521 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5581 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5641 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5701 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5761 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5821 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5881 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5941 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6001 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6061 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6121 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6181 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6241 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6301 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6361 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6421 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6481 cgaaaagtgc cacctgacgt c
//
