LOCUS       sequence-337774-        6923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Hook1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6923)
  AUTHORS   Feng Z, Yu CH
  TITLE     PI(3,4)P2-mediated membrane tubulation promotes integrin trafficking
            and invasive cell migration.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 May 11;118(19). pii: 2017645118. doi:
            10.1073/pnas.2017645118.
  PUBMED    33947811
REFERENCE   2  (bases 1 to 6923)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-05-11"; volume: "118(19). pii: 
            2017645118. doi"; pages: " 10.1073/pnas.2017645118"
COMMENT     Sequence Label: Hook1-EGFP
FEATURES             Location/Qualifiers
     source          1..6923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2863..2872
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2869..2871,2872..2874,2875..3588)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2869..2871,2872..2874,2875..3585)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2914..2935)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3175..3194)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3522..3543
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3711..3832
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3748..3767)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3802..3821
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3839..4294)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3976..3997)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4188..4207
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4321..4425
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4427..4784
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4427..4618
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4450..4470)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4635..4770
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4819..5613
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4819..5613
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4873..4892)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5483..5502
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5801..5820)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5845..5892
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6221..6809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6710..6729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttatg gaggagacgc agccgccgcc gcagcctaag
      661 ctgcccctgt gcgacagcct catgatctgg ctgcagacat tcaatactgc ctcaccttgt
      721 caagatgtca aacagctgac tagtggagtt gccatggcac aagttcttca tcaaattgat
      781 gcagcttggt ttaacgaatc ttggttaagc cgaattaaag aggatgttgg ggacaactgg
      841 agaataaagg ccagtaatgt aaagaaggtc cttcaaggaa ttatgagtta ttatcatgag
      901 tttttggggc agcagatttc agaagcactt atccctgatt taaaccaaat aaccgaatgt
      961 tcagatccag tggagcttgg gaggttgctc cagcttattt taggttgtgc gatcaactgt
     1021 gaaaagaagc aagaacatat tcaaaatata atgacactgg aagagtctgt tcaacatgtg
     1081 gtcatgactg ctattcaaga gttgatgagt aaagaaatat tgagctctcc tccaaatgat
     1141 gctgttggag aattggagca acagcttaaa agagccttgg aagaacttca ggaagcacta
     1201 gcagaaaaag aagagctgag gcaaagatgt gaagaattgg atatgcaggt gactacactt
     1261 caagatgaaa agaattcact ggtttctgaa aatgagatga tgaatgaaaa acttgaccag
     1321 ttggatggct cttttgatga tccaaacaca gtggttgcaa aaaagtattt tcatgcacaa
     1381 ttacaactag aacaattaca ggaagaaaac ttcaggcttg aagctgcaaa agatgattac
     1441 cgtgttcact gtgaagaact tgaaaagcag ctaatcgaat tccagcatag gaatgatgaa
     1501 ttgactagtc ttgcagaaga aacaagagcc ctgaaagatg aaatagatgt tcttagggct
     1561 acctctgata aagcaaataa actggagtca acagttgaga tatatcgtca gaagctacaa
     1621 gatctgaatg accttcgcaa gcaggtgaaa actttacagg aaaccaacat gatgtatatg
     1681 cataatacag tcagcttaga agaagaatta aaaaaagcaa atgcagcacg tacacaatta
     1741 gaaacataca aaaggcaggt tcaagatctt catgttaaac tttcctccga atccaagagg
     1801 gcagacacac tagcgtttga aatgaagcgg cttgaagaaa aacatgaagc tttacttaag
     1861 gaaaaagaga gactaattga gcagcgtgat actttgaaag aaacaaatga agagcttcga
     1921 tgttcacaag tacaacagga ccacctaaac caaacagatg catctgctac aaaaagttat
     1981 gagaatcttg ctgctgagat tatgccagtg gaatataggg aggtgtttat tcgactgcaa
     2041 catgaaaata agatgcttcg cttacagcaa gaaggctctg agaatgaacg tattgaggaa
     2101 cttcaggagc agctagaaca gaaacaccgt aaaatgaatg aactggaaac tgagcagagg
     2161 ctgagcaaag agcgtattag agaattgcag cagcagattg aggacctcca gaaatcttta
     2221 caggaacaag gttccaagtc tgaaggcgaa agttccagca aattaaagca gaagttggaa
     2281 gctcatatgg aaaaactcac agaggtccat gaagaattac agaagaaaca agaactcatt
     2341 gaagatcttc agccagatat aaatcaaaat gtacaaaaga tcaatgaact tgaagctgct
     2401 cttcagaaga aagatgaaga tatgaaagca atggaggaaa gatataaaat gtacttggag
     2461 aaagccagaa atgtaataaa aactttggat cccaagttaa atccagcatc agctgaaata
     2521 atgctactaa gaaagcagtt ggcagagaaa gagagaagaa ttgagattct ggagagtgaa
     2581 tgcaaagtag caaaattccg tgattatgaa gaaaaactca ttgtttctgc gtggtataat
     2641 aagagtctag cattccagaa actggggatg gaatctagac ttgtgagcgg cggtggtgcc
     2701 tgcagtgaca ctggtgcgtg cactcctgcg cggtctttct tagcgcagca acggcacatc
     2761 accaacacca gaagaaatct ctctgttaaa gtccctgcta caacatctga taagcttcga
     2821 attctgcagt cgacggtacc gcgggcccgg gatccaccgg tcgccaccat ggtgagcaag
     2881 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2941 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     3001 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     3061 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     3121 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     3181 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     3241 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     3301 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     3361 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     3421 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     3481 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     3541 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac
     3601 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3661 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3721 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3781 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     3841 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3901 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3961 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4021 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4081 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4141 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4201 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4261 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     4321 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4381 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     4441 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4501 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4561 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4621 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4681 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4741 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     4801 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4861 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4921 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4981 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     5041 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5101 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5161 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5221 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5281 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5341 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5401 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5461 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5521 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5581 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5641 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5701 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5761 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     5821 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     5881 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     5941 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     6001 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     6061 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6121 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6181 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6241 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6301 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6361 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6421 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6481 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6541 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6601 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6661 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6721 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6781 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6841 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6901 ataaccgtat taccgccatg cat
//
