LOCUS       sequence-257514-        5262 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 5262)
  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 5262)
  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: pEGFP-C1 hSlp4-a C2B
FEATURES             Location/Qualifiers
     source          1..5262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(671..673,674..676,677..1387)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(671..673,674..676,677..1387)
                     /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(716..737)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(977..996)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1324..1345
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2108..2229
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2145..2164)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2199..2218
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2236..2691)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2373..2394)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2585..2604
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2718..2822
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2824..3181
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2824..3015
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2847..2867)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3032..3167
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3216..4010
                     /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             3216..4010
                     /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(3270..3289)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3880..3899
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4198..4217)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4242..4289
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4618..5206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5107..5126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      721 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      781 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      841 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      901 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      961 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1021 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1081 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     1141 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1201 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1261 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     1321 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1381 gtacaagtcc ggactcagat ctcgagctca agcttcgaat tcttccccga ctggcttgcc
     1441 atcacacaaa ggcgagttgg tggtttcatt gaaatacatc ccagcctcca aaacccctgt
     1501 tggaggtgac cggaaaaaga gtaaaggtgg ggaaggggga gagctccagg tgtggatcaa
     1561 agaagccaag aacttgacgg ctgccaaagc aggagggact tcagacagct ttgtcaaggg
     1621 atacctcctt cccatgagga acaaggccag taaacgtaaa actcctgtga tgaagaagac
     1681 cctgaatcct cactacaacc atacatttgt ctacaatggt gtgaggctgg aagatctaca
     1741 gcatatgtgc ctggaactga ctgtgtggga ccgggagccc ctggccagca atgacttcct
     1801 gggaggggtc aggctgggtg ttggcactgg gatcagtaat ggggaagtgg tggactggat
     1861 ggactcgact ggggaagaag tgagcctgtg gcagaagatg cgacagtacc cagggtcttg
     1921 ggcagaaggg actctgcagc tccgttcctc aatggccaag cagaagctgg gtttatgagg
     1981 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     2041 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2101 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2161 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2221 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2281 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2341 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2401 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2461 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2521 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2581 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2641 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     2701 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2761 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2821 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2881 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2941 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3001 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3061 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3121 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3181 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3241 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3301 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3361 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     3421 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3481 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3541 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3601 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3661 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3721 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     3781 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3841 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3901 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3961 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4021 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4081 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4141 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     4201 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4261 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4321 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4381 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4441 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     4501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5221 ggccttttta cggttcctgg ccttttgctg gccttttgct ca
//
