LOCUS       sequence-305769-        7429 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 7429)
  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 7429)
  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 hSlp2-a
FEATURES             Location/Qualifiers
     source          1..7429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      548..557
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(554..556,557..559,560..1270)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(599..620)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(860..879)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1207..1228
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4158..4279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4195..4214)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4249..4268
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4286..4741)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4423..4444)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4635..4654
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4768..4872
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4874..5231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4874..5065
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4897..4917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5082..5217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5266..6060
                     /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             5266..6060
                     /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(5320..5339)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5930..5949
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6248..6267)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6292..6339
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6668..7256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7157..7176
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      541 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      601 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      661 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
      721 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
      781 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      841 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      901 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      961 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1021 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1081 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1141 gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg
     1201 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1261 gctgtacaag tccggactca gatctcgagc gattgactta agcttcctga ctgaagagga
     1321 acaagaggcc atcatgaagg ttttgcagcg ggatgctgct ctgaagaggg ccgaagaaga
     1381 gagagtcaga catttgcctg aaaaaattaa ggatgaccag cagctgaaga atatgagtgg
     1441 ccaatggttt tatgaagcca aggcaaaaag gcacagggac aaaatccatg gcgcagatat
     1501 catcagagca tctatgagaa agaagaggcc ccagatagca gctgagcaga gtaaagacag
     1561 agaaaatggg gcaaaggaaa gctgggtgaa taatgtcaac aaagatgctt tccttcctcc
     1621 agagctggct ggcgttgtag aagagccaga agaggatgca gcaccagcaa gcccgagttc
     1681 cagtgtggta aatccagctt ccagtgtgat tgatatgtcc caggaaaaca caaggaaacc
     1741 aaatgtgtct ccagagaagc agaggaagaa tccgtttaat agctccaagt tgccagaagg
     1801 tcactcgtca caacaaacta aaaatgaaca gtcaaaaaat ggaagaactg gtttatttca
     1861 gacttcaaaa gaggatgaat tgtcagagtc aaaagaaaag tcaactgtcg cagatacttc
     1921 aatccaaaag ttagagaaat caaagcagac tttgccaggc ctttcaaatg ggtcccaaat
     1981 caaggctcca atccccaaag ccaggaagat gatctacaaa tcaactgatt taaacaaaga
     2041 tgataaccag tcttttccta gacaaaggac agactccctg aaagcgagag gggctccgag
     2101 agggatcctc aagcgcaact ccagttccag tagcacagac tcagaaaccc ttcgttataa
     2161 tcacaacttt gaacccaaaa gcaaaattgt gtcacctggc ctaaccatcc atgagagaat
     2221 ttctgagaag gagcattctt tagaagacaa ctcttcccca aactccctgg agccattaaa
     2281 gcatgtgaga ttctctgcag tgaaggatga gcttccacag agtcctgggc taatccatgg
     2341 tcgggaagta ggagaattta gtgttttaga atctgacaga ttgaaaaatg gaatggaaga
     2401 tgcaggggac acagaagagt ttcagagtga ccctaagcct tctcaataca gaaagccttc
     2461 gctttttcat caatcaacct caagcccata tgtatcaaaa agtgaaacac atcagccaat
     2521 gacttctggt tcttttccaa ttaatgggct gcattctcat tcagaagttt taactgcaag
     2581 accacagtct atggagaatt caccaaccat caatgaaccc aaagataaat catcagaatt
     2641 aacaaggctt gaatctgtat tacccagaag ccctgctgat gaactgtctc attgtgttga
     2701 gcctgagcca tctcaggtgc caggtggcag ttctagagac cgtcagcaag gttcagaaga
     2761 agaacccagt cctgttttga aaactttgga aaggagtgcc gctaggaaaa tgccttccaa
     2821 aagtctagaa gacatttcat cagattcatc aaatcaagca aaagtagata atcagccaga
     2881 agaattagtg cgtagtgctg aagatgatga gaaaccagat cagaagccag ttacaaatga
     2941 atgcgtacca agaatttcca cagtgcctac acaacctgat aatccatttt ctcaccctga
     3001 caaactcaaa aggatgagca agtctgttcc agcatttctc caagatgagg tgagtggcag
     3061 tgtgatgagt gtttatagtg gagactttgg caatctggaa gttaaaggaa atattcagtt
     3121 tgcaattgaa tatgtggagt cactgaagga gttgcatgtt tttgtggccc agtgtaagga
     3181 cttagcagca gcggatgtaa aaaaacagcg ttcagaccca tatgtaaagg cctatttgct
     3241 accagacaaa ggcaaaatgg gcaagaagaa aacactcgta gtgaagaaaa ccttgaatcc
     3301 tgtgtataac gaaatactgc ggtataaaat tgaaaaacaa atcttaaaga cacagaaatt
     3361 gaacctgtcc atttggcatc gggatacatt taagcgcaat agtttcctag gggaggtgga
     3421 acttgatttg gaaacatggg actgggataa caaacagaat aaacaattga gatggtaccc
     3481 tctgaagcgg aagacagcac cagttgccct tgaagcagaa aacagaggtg aaatgaaact
     3541 agctctccag tatgtcccag agccagtccc tggtaaaaag cttcctacaa ctggagaagt
     3601 gcacatctgg gtgaaggaat gccttgatct accactgcta aggggaagtc atctaaattc
     3661 ttttgttaaa tgtaccatcc ttccagatac aagtaggaaa agtcgccaga agacaagagc
     3721 tgtagggaaa accaccaacc ctatcttcaa ccacactatg gtgtatgatg ggttcaggcc
     3781 tgaagatctg atggaagcct gtgtagagct tactgtctgg gaccattaca aattaaccaa
     3841 ccaatttttg ggaggtcttc gtattggctt tggaacaggt aaaagttatg ggactgaagt
     3901 ggactggatg gactctactt cagaggaagt tgctctctgg gagaagatgg taaactcccc
     3961 caatacttgg attgaagcaa cactgcctct cagaatgctt ttgattgcca agatttccaa
     4021 atgacccggg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     4081 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     4141 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4201 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4261 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     4321 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4381 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4441 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4501 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4561 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4621 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4681 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4741 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4801 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4861 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     4921 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4981 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5041 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5101 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5161 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5221 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5281 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5341 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5401 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     5461 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5521 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5581 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5641 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5701 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5761 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     5821 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5881 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5941 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6001 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6061 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     6121 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6181 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     6241 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     6301 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     6361 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     6421 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     6481 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     6541 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6601 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6661 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6721 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6781 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6841 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6901 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6961 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7021 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7081 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7141 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7201 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7261 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7321 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     7381 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttcc
//
