LOCUS       sequence-370126-        7672 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Retroviral expression of Polo-Like Kinase 1 (Plk1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7672)
  TITLE     Cheeseman Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7672)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pIC195
FEATURES             Location/Qualifiers
     source          1..7672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             117..533
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     463..485
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     501..517
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      568..577
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(574..576,577..579,580..1290)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(619..640)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(880..899)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1227..1248
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1336..1356
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1369..1413
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3432..3452)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3437..3766
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3617..3752
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3774..4172
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4341..4810
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4940..4957)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5111..5699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5191..5210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5859..6650,6651..6719))
                     /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     6482..6501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6720..6824)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             6855..7324
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    7389..7588
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca
       61 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
      121 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
      181 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
      241 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
      301 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
      361 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
      421 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
      481 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggccgga
      541 tcccagtgtg gtggtacgct accggtcgcc accatggtga gcaagggcga ggagctgttc
      601 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      661 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
      721 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
      781 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      841 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
      901 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
      961 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1021 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1081 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1141 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     1201 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1261 atcactctcg gcatggacga gctgtacaag tccggagggg atcggtggag ctccaccggt
     1321 ggcggccgct ctagagagaa tctttatttt cagggcgccg ccaaattcaa agaaaccgct
     1381 gctgctaaat tcgaacgcca gcacatggac agcggaggtg gaggtactag tctcgagatg
     1441 agtgctgcag tgactgcagg gaagctggca cgggcaccgg ccgaccctgg gaaagccggg
     1501 gtccccggag ttgcagctcc cggagctccg gcggcggctc caccggcgaa agagatcccg
     1561 gaggtcctag tggacccacg cagccggcgg cgctatgtgc ggggccgctt tttgggcaag
     1621 ggcggctttg ccaagtgctt cgagatctcg gacgcggaca ccaaggaggt gttcgcgggc
     1681 aagattgtgc ctaagtctct gctgctcaag ccgcaccaga gggagaagat gtccatggaa
     1741 atatccattc accgcagcct cgcccaccag cacgtcgtag gattccacgg ctttttcgag
     1801 gacaacgact tcgtgttcgt ggtgttggag ctctgccgcc ggaggtctct cctggagctg
     1861 cacaagagga ggaaagccct gactgagcct gaggcccgat actacctacg gcaaattgtg
     1921 cttggctgcc agtacctgca ccgaaaccga gttattcatc gagacctcaa gctgggcaac
     1981 cttttcctga atgaagatct ggaggtgaaa ataggggatt ttggactggc aaccaaagtc
     2041 gaatatgacg gggagaggaa gaagaccctg tgtgggactc ctaattacat agctcccgag
     2101 gtgctgagca agaaagggca cagtttcgag gtggatgtgt ggtccattgg gtgtatcatg
     2161 tataccttgt tagtgggcaa accacctttt gagacttctt gcctaaaaga gacctacctc
     2221 cggatcaaga agaatgaata cagtattccc aagcacatca accccgtggc cgcctccctc
     2281 atccagaaga tgcttcagac agatcccact gcccgcccaa ccattaacga gctgcttaat
     2341 gacgagttct ttacttctgg ctatatccct gcccgtctcc ccatcacctg cctgaccatt
     2401 ccaccaaggt tttcgattgc tcccagcagc ctggacccca gcaaccggaa gcccctcaca
     2461 gtcctcaata aaggcttgga gaaccccctg cctgagcgtc cccgggaaaa agaagaacca
     2521 gtggttcgag agacaggtga ggtggtcgac tgccacctca gtgacatgct gcagcagctg
     2581 cacagtgtca atgcctccaa gccctcggag cgtgggctgg tcaggcaaga ggaggctgag
     2641 gatcctgcct gcatccccat cttctgggtc agcaagtggg tggactattc ggacaagtac
     2701 ggccttgggt atcagctctg tgataacagc gtgggggtgc tcttcaatga ctcaacacgc
     2761 ctcatcctct acaatgatgg tgacagcctg cagtacatag agcgtgacgg cactgagtcc
     2821 tacctcaccg tgagttccca tcccaactcc ttgatgaaga agatcaccct ccttaaatat
     2881 ttccgcaatt acatgagcga gcacttgctg aaggcaggtg ccaacatcac gccgcgcgaa
     2941 ggtgatgagc tcgcccggct gccctaccta cggacctggt tccgcacccg cagcgccatc
     3001 atcctgcacc tcagcaacgg cagcgtgcag atcaacttct tccaggatca caccaagctc
     3061 atcttgtgcc cactgatggc agccgtgacc tacatcgacg agaagcggga cttccgcaca
     3121 taccgcctga gtctcctgga ggagtacggc tgctgcaagg agctggccag ccggctccgc
     3181 tacgcccgca ctatggtgga caagctgctg agctcacgct cggccagcaa ccgtctcaag
     3241 gcctcctaaa agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggat
     3301 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     3361 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattcgc cagcacagtg
     3421 gtcgaccctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3481 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3541 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3601 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3661 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3721 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttcatggcca
     3781 agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca
     3841 tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct
     3901 tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg
     3961 gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga
     4021 acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg
     4081 ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat
     4141 tgctgccctc tggttatgtg tgggagggct aagatcgata aaataaaaga ttttatttag
     4201 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     4261 aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt tcagatcaag
     4321 gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     4381 ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc
     4441 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     4501 cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     4561 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     4621 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc
     4681 gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
     4741 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
     4801 gggtctttca catgcagcat gtatcaaaat taatttggtt ttttttctta agtatttaca
     4861 ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4921 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4981 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5041 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5101 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5161 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5221 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5281 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5341 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5401 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5461 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5521 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5581 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5641 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5701 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5761 ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg cggccgcaaa
     5821 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5881 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5941 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6001 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6061 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6121 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6181 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6241 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6301 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6361 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6421 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6481 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6541 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6601 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6661 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6721 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6781 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6841 gtgccacctg cagcctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     6901 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     6961 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     7021 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     7081 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     7141 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     7201 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     7261 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     7321 ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc gacccaccac
     7381 cgggaggcaa gctggccagc aacttatctg tgtctgtccg attgtctagt gtctatgact
     7441 gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg gcggacccgt
     7501 ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt
     7561 gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca
     7621 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gt
//
