LOCUS       sequence-47764-e        8468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8468)
  AUTHORS   Gierke S, Wittmann T
  TITLE     EB1-Recruited Microtubule +TIP Complexes Coordinate Protrusion 
            Dynamics during 3D Epithelial Remodeling.
  JOURNAL   Curr Biol. 2012 May 8;22(9):753-62. Epub 2012 Apr 5.
  PUBMED    22483942
REFERENCE   2  (bases 1 to 8468)
  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: "Curr 
            Biol."; date: "2012-05-8"; volume: "22(9)"; pages: "753-62. Epub 
            2012 Apr 5"
COMMENT     Sequence Label: pShuttle-mCherry-EB1-C
FEATURES             Location/Qualifiers
     source          1..8468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..103
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     191..341
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
     enhancer        399..702
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        703..906
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     857..877
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      946..955
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             952..1659
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1097..1115)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1365..1384
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2116..2237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2153..2172)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2207..2226
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5428..5530
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     primer_bind     complement(5619..5636)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5790..6378)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5870..5889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7210..8004
                     /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             7210..8004
                     /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(7264..7283)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7874..7893
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(8052..8431)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8114..8135)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8326..8345
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 catcatcaat aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt
       61 ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt
      121 gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg
      181 gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag
      241 taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga
      301 agtgaaatct gaataatttt gtgttactca tagcgcgtaa tactgtaata gtaatcaatt
      361 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      421 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      481 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      541 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      601 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      661 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      721 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      781 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      841 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      901 agagctggtt tagtgaaccg tcagatccgc tagagatctg gtaccgccac catggtgagc
      961 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     1021 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     1081 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     1141 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     1201 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     1261 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     1321 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     1381 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     1441 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     1501 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     1561 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     1621 ggccgccact ccaccggcgg catggacgag ctgtacaagc tcgagtcaac acagagaacc
     1681 gctgcggctc ctaaggctgg ccctggtgtg gtgcgaaaga accctggtgt gggcaacgga
     1741 gacgacgagg cagctgagtt gatgcagcag gtcaacgtat tgaaacttac tgttgaagac
     1801 ttggagaaag agagggattt ctacttcgga aagctacgga acattgaatt gatttgccag
     1861 gagaacgagg gggaaaacga ccctgtattg cagaggattg tagacattct gtatgccaca
     1921 gatgaaggct ttgtgatacc tgatgaaggg ggcccacagg aggagcaaga agagtattaa
     1981 gatatccgat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
     2041 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     2101 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2161 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2221 ctcatcaatg tatcttaacg cggatctggg cgtggttaag ggtgggaaag aatatataag
     2281 gtgggggtct tatgtagttt tgtatctgtt ttgcagcagc cgccgccgcc atgagcacca
     2341 actcgtttga tggaagcatt gtgagctcat atttgacaac gcgcatgccc ccatgggccg
     2401 gggtgcgtca gaatgtgatg ggctccagca ttgatggtcg ccccgtcctg cccgcaaact
     2461 ctactacctt gacctacgag accgtgtctg gaacgccgtt ggagactgca gcctccgccg
     2521 ccgcttcagc cgctgcagcc accgcccgcg ggattgtgac tgactttgct ttcctgagcc
     2581 cgcttgcaag cagtgcagct tcccgttcat ccgcccgcga tgacaagttg acggctcttt
     2641 tggcacaatt ggattctttg acccgggaac ttaatgtcgt ttctcagcag ctgttggatc
     2701 tgcgccagca ggtttctgcc ctgaaggctt cctcccctcc caatgcggtt taaaacataa
     2761 ataaaaaacc agactctgtt tggatttgga tcaagcaagt gtcttgctgt ctttatttag
     2821 gggttttgcg cgcgcggtag gcccgggacc agcggtctcg gtcgttgagg gtcctgtgta
     2881 ttttttccag gacgtggtaa aggtgactct ggatgttcag atacatgggc ataagcccgt
     2941 ctctggggtg gaggtagcac cactgcagag cttcatgctg cggggtggtg ttgtagatga
     3001 tccagtcgta gcaggagcgc tgggcgtggt gcctaaaaat gtctttcagt agcaagctga
     3061 ttgccagggg caggcccttg gtgtaagtgt ttacaaagcg gttaagctgg gatgggtgca
     3121 tacgtgggga tatgagatgc atcttggact gtatttttag gttggctatg ttcccagcca
     3181 tatccctccg gggattcatg ttgtgcagaa ccaccagcac agtgtatccg gtgcacttgg
     3241 gaaatttgtc atgtagctta gaaggaaatg cgtggaagaa cttggagacg cccttgtgac
     3301 ctccaagatt ttccatgcat tcgtccataa tgatggcaat gggcccacgg gcggcggcct
     3361 gggcgaagat atttctggga tcactaacgt catagttgtg ttccaggatg agatcgtcat
     3421 aggccatttt tacaaagcgc gggcggaggg tgccagactg cggtataatg gttccatccg
     3481 gcccaggggc gtagttaccc tcacagattt gcatttccca cgctttgagt tcagatgggg
     3541 ggatcatgtc tacctgcggg gcgatgaaga aaacggtttc cggggtaggg gagatcagct
     3601 gggaagaaag caggttcctg agcagctgcg acttaccgca gccggtgggc ccgtaaatca
     3661 cacctattac cggctgcaac tggtagttaa gagagctgca gctgccgtca tccctgagca
     3721 ggggggccac ttcgttaagc atgtccctga ctcgcatgtt ttccctgacc aaatccgcca
     3781 gaaggcgctc gccgcccagc gatagcagtt cttgcaagga agcaaagttt ttcaacggtt
     3841 tgagaccgtc cgccgtaggc atgcttttga gcgtttgacc aagcagttcc aggcggtccc
     3901 acagctcggt cacctgctct acggcatctc gatccagcat atctcctcgt ttcgcgggtt
     3961 ggggcggctt tcgctgtacg gcagtagtcg gtgctcgtcc agacgggcca gggtcatgtc
     4021 tttccacggg cgcagggtcc tcgtcagcgt agtctgggtc acggtgaagg ggtgcgctcc
     4081 gggctgcgcg ctggccaggg tgcgcttgag gctggtcctg ctggtgctga agcgctgccg
     4141 gtcttcgccc tgcgcgtcgg ccaggtagca tttgaccatg gtgtcatagt ccagcccctc
     4201 cgcggcgtgg cccttggcgc gcagcttgcc cttggaggag gcgccgcacg aggggcagtg
     4261 cagacttttg agggcgtaga gcttgggcgc gagaaatacc gattccgggg agtaggcatc
     4321 cgcgccgcag gccccgcaga cggtctcgca ttccacgagc caggtgagct ctggccgttc
     4381 ggggtcaaaa accaggtttc ccccatgctt tttgatgcgt ttcttacctc tggtttccat
     4441 gagccggtgt ccacgctcgg tgacgaaaag gctgtccgtg tccccgtata cagacttgag
     4501 agggagttta aacgaattca atagcttgtt gcatgggcgg cgatataaaa tgcaaggtgc
     4561 tgctcaaaaa atcaggcaaa gcctcgcgca aaaaagaaag cacatcgtag tcatgctcat
     4621 gcagataaag gcaggtaagc tccggaacca ccacagaaaa agacaccatt tttctctcaa
     4681 acatgtctgc gggtttctgc ataaacacaa aataaaataa caaaaaaaca tttaaacatt
     4741 agaagcctgt cttacaacag gaaaaacaac ccttataagc ataagacgga ctacggccat
     4801 gccggcgtga ccgtaaaaaa actggtcacc gtgattaaaa agcaccaccg acagctcctc
     4861 ggtcatgtcc ggagtcataa tgtaagactc ggtaaacaca tcaggttgat tcacatcggt
     4921 cagtgctaaa aagcgaccga aatagcccgg gggaatacat acccgcaggc gtagagacaa
     4981 cattacagcc cccataggag gtataacaaa attaatagga gagaaaaaca cataaacacc
     5041 tgaaaaaccc tcctgcctag gcaaaatagc accctcccgc tccagaacaa catacagcgc
     5101 ttccacagcg gcagccataa cagtcagcct taccagtaaa aaagaaaacc tattaaaaaa
     5161 acaccactcg acacggcacc agctcaatca gtcacagtgt aaaaaagggc caagtgcaga
     5221 gcgagtatat ataggactaa aaaatgacgt aacggttaaa gtccacaaaa aacacccaga
     5281 aaaccgcacg cgaacctacg cccagaaacg aaagccaaaa aacccacaac ttcctcaaat
     5341 cgtcacttcc gttttcccac gttacgtcac ttcccatttt aagaaaacta caattcccaa
     5401 cacatacaag ttactccgcc ctaaaaccta cgtcacccgc cccgttccca cgccccgcgc
     5461 cacgtcacaa actccacccc ctcattatca tattggcttc aatccaaaat aaggtatatt
     5521 attgatgatg ttaattaaca tgcatggatc catatgcggt gtgaaatacc gcacagatgc
     5581 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5641 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5701 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5761 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5821 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5881 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5941 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6001 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6061 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6121 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6181 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6241 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6301 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6361 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6421 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6481 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6541 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6601 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6661 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6721 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6781 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6841 ttgcgcaacg ttgttgccat tgctgcagcc atgagattat caaaaaggat cttcacctag
     6901 atccttttca cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc
     6961 tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt
     7021 gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg
     7081 ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc
     7141 ttgccgccaa ggatctgatg gcgcagggga tcaagctctg atcaagagac aggatgagga
     7201 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     7261 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     7321 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     7381 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     7441 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     7501 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     7561 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     7621 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     7681 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     7741 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     7801 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     7861 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     7921 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     7981 cgccttcttg acgagttctt ctgaattttg ttaaaatttt tgttaaatca gctcattttt
     8041 taaccaatag gccgaaatcg gcaccatccc ttataaatca aaagaataga ccgagatagg
     8101 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     8161 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     8221 aagttttttg tggtcgaggt gccgtaaaga tctaaatcgg aaccctaaag ggagcccccg
     8281 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     8341 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     8401 cgcgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggatc gaattaattc
     8461 ttaattaa
//
