LOCUS       sequence-337700-        5610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AKTIP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5610)
  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 5610)
  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: mRuby2-AKTIP
FEATURES             Location/Qualifiers
     source          1..5610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        74..377
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        378..581
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     532..552
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      620..629
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             626..1336
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     polyA_signal    2411..2532
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2448..2467)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2502..2521
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2539..2994)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2676..2697)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2888..2907
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3021..3125
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3127..3484
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3127..3318
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3150..3170)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3335..3470
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3519..4313
                     /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             3519..4313
                     /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(3573..3592)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4183..4202
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4501..4520)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4545..4592
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4921..5509
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5410..5429
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
       61 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      121 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      181 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      241 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      301 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      361 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      421 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      481 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      541 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      601 tccgctagcg ctaccggtcg ccaccatggt gtctaagggc gaagagctga tcaaggaaaa
      661 tatgcgtatg aaggtggtca tggaaggttc ggtcaacggc caccaattca aatgcacagg
      721 tgaaggagaa ggcaatccgt acatgggaac tcaaaccatg aggatcaaag tcatcgaggg
      781 aggacccctg ccatttgcct ttgacattct tgccacgtcg ttcatgtatg gcagccgtac
      841 ttttatcaag tacccgaaag gcattcctga tttctttaaa cagtcctttc ctgagggttt
      901 tacttgggaa agagttacga gatacgaaga tggtggagtc gtcaccgtca tgcaggacac
      961 cagccttgag gatggctgtc tcgtttacca cgtccaagtc agaggggtaa actttccctc
     1021 caatggtccc gtgatgcaga agaagaccaa gggttgggag cctaatacag agatgatgta
     1081 tccagcagat ggtggtctga ggggatacac tcatatggca ctgaaagttg atggtggtgg
     1141 ccatctgtct tgctctttcg taacaactta caggtcaaaa aagaccgtcg ggaacatcaa
     1201 gatgcccggt atccatgccg ttgatcaccg cctggaaagg ttagaggaaa gtgacaatga
     1261 aatgttcgta gtacaacgcg aacacgcagt tgccaagttc gccgggcttg gtggtgggat
     1321 ggacgagctg tacaagtccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga
     1381 catgaaccct ttctggagca tgtctacaag ctctgtacgc aaacgatctg aaggtgaaga
     1441 gaagacatta acaggggacg tgaaaaccag tcctccacga actgcaccaa agaaacagct
     1501 gccttctatt cccaaaaatg ctttgcccat aactaagcct acatctcctg ccccagcagc
     1561 acagtcaaca aatggcacgc atgcgtccta tggacccttc tacctggaat actctcttct
     1621 tgcagaattt accttggttg tgaagcagaa gctaccaggc gtctatgtgc agccatctta
     1681 tcgctctgca ttaatgtggt ttggagtaat attcatacgg catggacttt accaagatgg
     1741 cgtatttaag tttacagttt acatccctga taactatcca gatggtgact gtccacgctt
     1801 ggtgttcgat attcctgtct ttcacccgct agttgatccc acctcaggtg agctggatgt
     1861 gaagagagca tttgcaaaat ggaggcggaa ccataatcat atttggcagg tattaatgta
     1921 tgcaaggaga gttttctaca agattgatac agcaagcccc ctgaacccag aggctgcagt
     1981 actgtatgaa aaagatattc agctttttaa aagtaaagtt gttgacagtg ttaaggtgtg
     2041 cactgctcgt ttgtttgacc aacctaaaat agaagacccc tatgcaatta gcttttctcc
     2101 atggaatcct tctgtacatg atgaagccag agaaaagatg ctgactcaga aaaagcctga
     2161 agaacagcac aataaaagtg ttcatgttgc tggcctgtca tgggtaaagc ctggctcagt
     2221 acagcctttc agtaaagaag agaaaacagt ggcgacttaa gtcgacggta ccgcgggccc
     2281 gggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     2341 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     2401 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2461 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2521 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2581 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2641 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2701 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2761 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2821 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2881 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2941 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3001 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3061 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3121 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3181 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3241 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3301 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3361 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3421 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3481 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3541 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3601 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3661 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     3721 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3781 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3841 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3901 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3961 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4021 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4081 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4141 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4201 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4261 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4321 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4381 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4441 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     4501 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4561 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4621 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4681 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4741 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     4801 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4861 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4921 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4981 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5041 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5101 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5161 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5221 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5281 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5341 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5401 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5461 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5521 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5581 gttatcccct gattctgtgg ataaccgtat
//
