LOCUS       sequence-446062-        6572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector with the YPet fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6572)
  AUTHORS   Conway JRW, Joshi O, Kaivola J, Follain G, Gounis M, Kuhl D, Ivaska 
            J
  TITLE     Dynamic regulation of integrin beta1 phosphorylation supports 
            invasion of breast cancer cells.
  JOURNAL   Nat Cell Biol. 2025 May 26. doi: 10.1038/s41556-025-01663-4.
  PUBMED    40419795
REFERENCE   2  (bases 1 to 6572)
  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 Cell 
            Biol. 2025 May 26. doi: 10.1038/s41556-025-01663-4."
COMMENT     Sequence Label: pEF.DEST51-YPet
FEATURES             Location/Qualifiers
     source          1..6572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(166..185)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        473..1651
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          704..1642
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1599..1619
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1668..1687
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1668..1686
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1718..1742
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1780..1789
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1786..1788,1789..1791,1792..2505)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLLCTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
                     EFLTAAGITEGMNELY*"
     primer_bind     complement(1831..1852)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2092..2111)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(2520..2544)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2570..2611
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2621..2638
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2661..2678)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2667..2891
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2937..3365
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3024..3043)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3234..3255
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3374..3394)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3379..3708
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3559..3694
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3621..3640
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3756..3803
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3822..4220
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4378..4499
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4415..4434)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4469..4488
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4548..4564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4548..4564)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4561..4583)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4572..4588
                     /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(4596..4602,4603..4620,4621..4626))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4641..4662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4779..4796)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4939..5527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5019..5038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5698..6489,6490..6558))
                     /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     6321..6340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6559..6572,1..91))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
       61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      121 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
      181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      361 ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga
      421 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg
      481 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
      541 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
      601 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
      661 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
      721 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
      781 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
      841 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
      901 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
      961 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     1021 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     1081 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     1141 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     1201 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     1261 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
     1321 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     1381 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     1441 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     1501 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     1561 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     1621 aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtactaa tacgactcac
     1681 tatagggaga cccaagctgg ctaggtaagc ttgatcaaca agtttgtaca aaaaagcagg
     1741 ctggcgccgg aaccaattca gtcgactgga tccggtaccg ccaccatggt gagcaagggc
     1801 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1861 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1921 aagcttctat gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1981 ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     2041 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2101 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2161 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2221 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     2281 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     2341 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     2401 tccgccctgt tcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcctg
     2461 accgccgccg ggatcactga gggcatgaac gagctgtact aatgactcga gatatctaga
     2521 cccagctttc ttgtacaaag tggttgatct agagggcccg cggttcgaag gtaagcctat
     2581 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg
     2641 agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2701 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2761 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2821 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     2881 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     2941 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3001 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3061 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3121 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3181 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3241 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3301 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3361 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3421 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3481 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3541 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3601 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3661 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3721 gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt
     3781 atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag cctttgtctc
     3841 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
     3901 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
     3961 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
     4021 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
     4081 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
     4141 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
     4201 gttatgtgtg ggagggctaa gcacttcgtg gccgaggagc aggactgaca cgtgctacga
     4261 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4321 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4381 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4441 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4501 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     4561 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4621 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4681 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4741 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4801 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4861 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagga accgtaaaaa
     4921 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4981 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5041 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5101 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5161 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5221 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5281 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5341 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5401 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5461 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5521 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5581 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5641 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5701 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5761 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5821 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5881 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5941 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6001 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6061 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6121 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6181 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6241 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6301 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6361 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6421 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6481 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6541 gaaatgttga atactcatac tcttcctttt tc
//
