LOCUS       sequence-139005-        7420 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 7420)
  AUTHORS   Galvez-Santisteban M, Rodriguez-Fraticelli AE, Bryant DM, 
            Vergarajauregui S, Yasuda T, Banon-Rodriguez I, Bernascone I, Datta 
            A, Spivak N, Young K, Slim CL, Brakeman PR, Fukuda M, Mostov KE, 
            Martin-Belmonte F
  TITLE     Synaptotagmin-like proteins control the formation of a single apical
            membrane domain in epithelial cells.
  JOURNAL   Nat Cell Biol. 2012 Jul 22. doi: 10.1038/ncb2541.
  PUBMED    22820376
REFERENCE   2  (bases 1 to 7420)
  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 Cell 
            Biol. 2012 Jul 22. doi: 10.1038/ncb2541."
COMMENT     Sequence Label: pmCherry-C1 hSlp2-a
FEATURES             Location/Qualifiers
     source          1..7420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             620..1327
                     /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(765..783)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1033..1052
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4215..4336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4252..4271)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4306..4325
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4343..4798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4480..4501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4692..4711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4825..4929
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4931..5288
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4931..5122
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4954..4974)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5139..5274
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5323..6117
                     /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             5323..6117
                     /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(5377..5396)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5987..6006
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6305..6324)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6349..6396
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6725..7313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7214..7233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
      661 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
      721 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
      781 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
      841 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
      901 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
      961 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     1021 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
     1081 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     1141 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     1201 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     1261 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     1321 gtacaagtcc ggactcagat ctcgagcgat tgacttaagc ttcctgactg aagaggaaca
     1381 agaggccatc atgaaggttt tgcagcggga tgctgctctg aagagggccg aagaagagag
     1441 agtcagacat ttgcctgaaa aaattaagga tgaccagcag ctgaagaata tgagtggcca
     1501 atggttttat gaagccaagg caaaaaggca cagggacaaa atccatggcg cagatatcat
     1561 cagagcatct atgagaaaga agaggcccca gatagcagct gagcagagta aagacagaga
     1621 aaatggggca aaggaaagct gggtgaataa tgtcaacaaa gatgctttcc ttcctccaga
     1681 gctggctggc gttgtagaag agccagaaga ggatgcagca ccagcaagcc cgagttccag
     1741 tgtggtaaat ccagcttcca gtgtgattga tatgtcccag gaaaacacaa ggaaaccaaa
     1801 tgtgtctcca gagaagcaga ggaagaatcc gtttaatagc tccaagttgc cagaaggtca
     1861 ctcgtcacaa caaactaaaa atgaacagtc aaaaaatgga agaactggtt tatttcagac
     1921 ttcaaaagag gatgaattgt cagagtcaaa agaaaagtca actgtcgcag atacttcaat
     1981 ccaaaagtta gagaaatcaa agcagacttt gccaggcctt tcaaatgggt cccaaatcaa
     2041 ggctccaatc cccaaagcca ggaagatgat ctacaaatca actgatttaa acaaagatga
     2101 taaccagtct tttcctagac aaaggacaga ctccctgaaa gcgagagggg ctccgagagg
     2161 gatcctcaag cgcaactcca gttccagtag cacagactca gaaacccttc gttataatca
     2221 caactttgaa cccaaaagca aaattgtgtc acctggccta accatccatg agagaatttc
     2281 tgagaaggag cattctttag aagacaactc ttccccaaac tccctggagc cattaaagca
     2341 tgtgagattc tctgcagtga aggatgagct tccacagagt cctgggctaa tccatggtcg
     2401 ggaagtagga gaatttagtg ttttagaatc tgacagattg aaaaatggaa tggaagatgc
     2461 aggggacaca gaagagtttc agagtgaccc taagccttct caatacagaa agccttcgct
     2521 ttttcatcaa tcaacctcaa gcccatatgt atcaaaaagt gaaacacatc agccaatgac
     2581 ttctggttct tttccaatta atgggctgca ttctcattca gaagttttaa ctgcaagacc
     2641 acagtctatg gagaattcac caaccatcaa tgaacccaaa gataaatcat cagaattaac
     2701 aaggcttgaa tctgtattac ccagaagccc tgctgatgaa ctgtctcatt gtgttgagcc
     2761 tgagccatct caggtgccag gtggcagttc tagagaccgt cagcaaggtt cagaagaaga
     2821 acccagtcct gttttgaaaa ctttggaaag gagtgccgct aggaaaatgc cttccaaaag
     2881 tctagaagac atttcatcag attcatcaaa tcaagcaaaa gtagataatc agccagaaga
     2941 attagtgcgt agtgctgaag atgatgagaa accagatcag aagccagtta caaatgaatg
     3001 cgtaccaaga atttccacag tgcctacaca acctgataat ccattttctc accctgacaa
     3061 actcaaaagg atgagcaagt ctgttccagc atttctccaa gatgaggtga gtggcagtgt
     3121 gatgagtgtt tatagtggag actttggcaa tctggaagtt aaaggaaata ttcagtttgc
     3181 aattgaatat gtggagtcac tgaaggagtt gcatgttttt gtggcccagt gtaaggactt
     3241 agcagcagcg gatgtaaaaa aacagcgttc agacccatat gtaaaggcct atttgctacc
     3301 agacaaaggc aaaatgggca agaagaaaac actcgtagtg aagaaaacct tgaatcctgt
     3361 gtataacgaa atactgcggt ataaaattga aaaacaaatc ttaaagacac agaaattgaa
     3421 cctgtccatt tggcatcggg atacatttaa gcgcaatagt ttcctagggg aggtggaact
     3481 tgatttggaa acatgggact gggataacaa acagaataaa caattgagat ggtaccctct
     3541 gaagcggaag acagcaccag ttgcccttga agcagaaaac agaggtgaaa tgaaactagc
     3601 tctccagtat gtcccagagc cagtccctgg taaaaagctt cctacaactg gagaagtgca
     3661 catctgggtg aaggaatgcc ttgatctacc actgctaagg ggaagtcatc taaattcttt
     3721 tgttaaatgt accatccttc cagatacaag taggaaaagt cgccagaaga caagagctgt
     3781 agggaaaacc accaacccta tcttcaacca cactatggtg tatgatgggt tcaggcctga
     3841 agatctgatg gaagcctgtg tagagcttac tgtctgggac cattacaaat taaccaacca
     3901 atttttggga ggtcttcgta ttggctttgg aacaggtaaa agttatggga ctgaagtgga
     3961 ctggatggac tctacttcag aggaagttgc tctctgggag aagatggtaa actcccccaa
     4021 tacttggatt gaagcaacac tgcctctcag aatgcttttg attgccaaga tttccaaatg
     4081 acccgggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     4141 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     4201 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4261 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4321 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     4381 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     4441 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     4501 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4561 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4621 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4681 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4741 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     4801 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4861 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4921 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4981 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5041 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5101 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5161 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5221 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5281 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5341 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5401 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5461 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     5521 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5581 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5641 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5701 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5761 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5821 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     5881 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5941 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     6001 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6061 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6121 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     6181 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     6241 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     6301 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     6361 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     6421 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     6481 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     6541 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     6601 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6661 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6721 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6781 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6841 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6901 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6961 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7021 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     7081 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7141 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7201 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     7261 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7321 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     7381 ctgcgttatc ccctgattct gtggataacc gtattaccgc
//
