LOCUS       sequence-313781-        7733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP tagged MLKP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7733)
  AUTHORS   Rathbun LI, Colicino EG, Manikas J, O'Connell J, Krishnan N, Reilly 
            NS, Coyne S, Erdemci-Tandogan G, Garrastegui A, Freshour J, Santra 
            P, Manning ML, Amack JD, Hehnly H
  TITLE     Cytokinetic bridge triggers de novo lumen formation in vivo.
  JOURNAL   Nat Commun. 2020 Mar 9;11(1):1269. doi: 10.1038/s41467-020-15002-8.
  PUBMED    32152267
REFERENCE   2  (bases 1 to 7733)
  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 
            Commun."; date: "2020-03-9"; volume: "11(1):1269. doi"; pages: " 
            10.1038/s41467-020-15002-8"
COMMENT     Sequence Label: pCS2-GFP-MKLP1
FEATURES             Location/Qualifiers
     source          1..7733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1064..1073
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1070..1072,1073..1075,1076..1786)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1070..1072,1073..1075,1076..1786)
                     /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(1115..1136)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1376..1395)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1723..1744
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4763..4897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4787..4806)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4841..4860
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5010..5030)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5010..5028)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5049..5065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5049..5065)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5062..5084)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5073..5089
                     /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(5097..5103,5104..5121,5122..5127))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5142..5163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5280..5297)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5451..6039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5531..5550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6210..7001,7002..7070))
                     /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     6833..6852
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7071..7175)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7201..7656)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7338..7359)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7550..7569
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcagccg gtcgccacca tggtgagcaa
     1081 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1141 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1201 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1261 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     1321 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1381 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1441 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1501 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     1561 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1621 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1681 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1741 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg gactcagatc
     1801 tcgagctcaa gcttcgaatt ctgcagtcga catgaagtca gcgagagcta ggacaccccg
     1861 gaaacctacc ttgaaaaaag ggtcccaaac gaaccttaaa gacccagttg gggtatactg
     1921 tagggtgcgc ccactgggct ttcctgatca agagtgttgc atagaagtga tcaataatac
     1981 aactgttcag cttcatactc ctgagggcta cagactcaac cgaaatggag actataagga
     2041 gactcagtat tcatttaaac aagtatttgg cactcacacc acccagaagg aactctttga
     2101 tgttgtggct aatcccttgg tcaatgacct cattcatggc aaaaatggtc ttctttttac
     2161 atatggtgtg acgggaagtg gaaaaactca cacaatgact ggttctccag aggaaggagg
     2221 gctgcttcct cgttgtttgg acatgatctt taacagtata gggtcatttc aagctaaacg
     2281 atatgttttc aaatctaatg ataggaatag tatggatata cagtgtgagg ttgatgcctt
     2341 attagaacgt cagaaaagag aagctatgcc caatccaaag acttcttcta gcaaacgaca
     2401 agtagatcca gagtttgcag atatgataac tgtacaagaa ttctgcaaag cagaagaggt
     2461 tgatgaagat agtgtctatg gtgtatttgt ctcttatatt gaaatatata ataattacat
     2521 atatgatcta ttggaagagg tgccgtttga tcccataaaa cccaaacctc cacaatctaa
     2581 attgcttcgt gaagataaga accataacat gtatgttgca ggatgtacag aagttgaagt
     2641 gaaatctact gaggaggctt ttgaagtttt ctggagaggc cagaaaaaga gacgtattgc
     2701 taatacccat ttgaatcgtg agtccagccg ttcccatagc gtgttcaaca ttaaattagt
     2761 tcaggctccc ttggatgcag atggagacaa tgtcttacag gaaaaagaac aaatcactat
     2821 aagtcagttg tccttggtag atcttgctgg aagtgaaaga actaaccgga ccagagcaga
     2881 agggaacaga ttacgtgaag ctggtaatat taatcagtca ctaatgacgc taagaacatg
     2941 tatggatgtc ctaagagagg accaaatgta tggaactaac aagatggttc catatcgaga
     3001 ttcaaagtta acccatctgt tcaagaacta ctttgatggg gaaggaaaag tgcggatgat
     3061 cgtgtgtgtg aaccccaagg ctgaagatta tgaagaaaac ttgcaagtca tgagatttgc
     3121 ggaagtgact caagaagttg aagtagcaag acctgtagac aaggcaatat gtggtttaac
     3181 gcctgggagg agatacagaa accagcctcg aggtccagtt ggaaatgaac cattggttac
     3241 tgacgtggtt ttgcagagtt ttccaccttt gccgtcatgc gaaattttgg atatcaacga
     3301 tgagcagaca cttccaaggc tgattgaagc cttagagaaa cgacataact tacgacaaat
     3361 gatgattgat gagtttaaca aacaatctaa tgcttttaaa gctttgttac aagaatttga
     3421 caatgctgtt ttaagtaaag aaaaccacat gcaagggaaa ctaaatgaaa aggagaagat
     3481 gatctcagga caggaattgg aaatagaacg actggaaaag aaaaacaaaa ctttagaata
     3541 taagattgag attttagaga aaacaactac tatctatgag gaagataaac gcaatttgca
     3601 acaggaactt gaaactcaga accagaaact tcagcgacag ttttctgaca aacgcagatt
     3661 agaagccagg ttgcaaggca tggtgacaga aacgacaatg aagtgggaga aagaatgtga
     3721 gcgtagagtg gcagccaaac agctggagat gcaggataaa ctctgggtta aagatgaaaa
     3781 gctgaaacaa ctgaaggcta ttgttactga acctaaaact gagaagccag agagaccctc
     3841 tcgggagcga gatcgagaaa aagttactca aagatctgtt tctccatcac ctgtgcctct
     3901 ttctagtaac tatattgctc agatttccaa cggccagcaa ctcatgagcc agccacagct
     3961 acataggcgc tctaactctt gcagcagcat ttctgtagct tcctgtattt cggaatggga
     4021 gctgaaaatt cctacgtaca acacacctct caaagtcaca tctattgcaa ggcgtaggca
     4081 gcaggagcca ggacaaagca aaacttgtat cgtgtcagac agaaggcgag ggatgtactg
     4141 gactgaaggc agggaggtgg ttcctacatt cagaaatgag atagaaatag aagaggatca
     4201 ttgcggcagg ttactctttc aacctgatca gaacgcacca ccaattcgtc tccgacacag
     4261 acgatcacgc tctgcaggag acagatgggt agatcataag cccgcctcta acatgcaaac
     4321 tgaaacagtc atgcagccac atgtccctca tgccatcaca gtatctgttg caaatgaaaa
     4381 ggcactagct aagtgtgaga agtacatgct gacccaccag gaactagcct ccgatgggga
     4441 gattgaaact aaactaatta agggtgatat ttataaaaca aggggtggtg gacaatctgt
     4501 tcagtttact gatattgaga ctttaaagca agaatcacca aatggtagtc gaaaacgaag
     4561 atcttccaca gtagcacctg cccaaccaga tggtgcagag tctgaatgga ccgatgtaga
     4621 aacaaggtgt tctgtggctg tggagatgag agcaggatcc cagctgggac ctggatatca
     4681 gcatcacgca caacccaagc gcaaaaagcc agtcgacggt accgcggctg cagtctagaa
     4741 ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca
     4801 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     4861 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     4921 tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg cggcgccaat
     4981 gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     5041 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5101 atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5161 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5221 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5281 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5341 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5401 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5461 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5521 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5581 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5641 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5701 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5761 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5821 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5881 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5941 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6001 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6061 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6121 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6181 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6241 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6301 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6361 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6421 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6481 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6541 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6601 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6661 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6721 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6781 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6841 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6901 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6961 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7021 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7081 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7141 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7201 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     7261 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     7321 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     7381 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     7441 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     7501 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     7561 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     7621 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     7681 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg acc
//
