LOCUS       sequence-122784-        7355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression, Bacterial Expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7355)
  AUTHORS   Giordano F, Saheki Y, Idevall-Hagren O, Colombo SF, Pirruccello M, 
            Milosevic I, Gracheva EO, Bagriantsev SN, Borgese N, De Camilli P
  TITLE     PI(4,5)P(2)-dependent and Ca(2+)-regulated ER-PM interactions 
            mediated by the extended synaptotagmins.
  JOURNAL   Cell. 2013 Jun 20;153(7):1494-509. doi: 10.1016/j.cell.2013.05.026.
  PUBMED    23791178
REFERENCE   2  (bases 1 to 7355)
  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"; doi: 
            "10.1016/j.cell.2013.05"; journalName: "Cell"; date: "2013-06-20- 
            20"; volume: "153"; issue: "7"; pages: "1494-509"
COMMENT     Sequence Label: Myc-E-Syt1
FEATURES             Location/Qualifiers
     source          1..7355
                     /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"
     polyA_signal    4150..4271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4187..4206)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4241..4260
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4278..4733)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4415..4436)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4627..4646
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4760..4864
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4866..5223
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4866..5057
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4889..4909)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5074..5209
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5258..6052
                     /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             5258..6052
                     /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(5312..5331)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5922..5941
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6240..6259)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6284..6331
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6660..7248
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7149..7168
                     /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 tgcagaagct gatctcagag gaggacctgt ccggactcag
      661 atctcgagct caagcttcga attctgcagt cgacatggag cgatctccag gagagggccc
      721 cagccccagc cccatggacc agccctctgc tccctccgac cccactgacc agccccccgc
      781 tgctcacgca aagccagacc caggttctgg gggccaacct gctggccctg gcgcggcggg
      841 tgaggccctg gcggtgctga cttcattcgg gaggcggttg ctggtgctga tacctgtgta
      901 tttggccggg gcagtgggac tcagcgtggg tttcgtgctc ttcggcctcg ccctctacct
      961 gggctggcgc cgggtccgcg acgagaaaga acggagcctt cgagcagcga ggcagctact
     1021 ggacgacgag gagcagctca ctgcgaaaac tctctatatg agtcatcgag agctacctgc
     1081 ctgggtcagc ttcccagacg tggaaaaggc tgaatggctc aataagattg tggcccaggt
     1141 ctggcccttc ctgggccagt atatggagaa gcttctggct gaaactgtgg ctccggctgt
     1201 taggggatct aacccccatc tgcaaacatt tacatttaca cgagtggaac tgggtgaaaa
     1261 gccattgcgc atcattggag tcaaggttca cccaggtcag agaaaagagc agatcctgct
     1321 ggacttgaac atcagctatg taggtgatgt gcagattgat gtggaagtga agaaatattt
     1381 ttgcaaagca ggagtcaagg gcatgcagct acatggcgtt ttgcgggtga tactggagcc
     1441 actcattggg gaccttccct tcgtgggggc tgtgtcaatg ttcttcatcc gacgcccgac
     1501 cctagacatc aactggacag ggatgaccaa cctgctggat atcccaggac ttagctcact
     1561 ctctgacacc atgatcatgg actccattgc tgccttcctc gtgttgccca accgattact
     1621 ggtgcccctt gtgcctgacc ttcaagatgt ggctcagttg cgttcccctc tgcccagggg
     1681 cattattcga attcacctgc tggctgctcg agggctgagt tccaaggaca aatatgtgaa
     1741 gggcctgatt gagggcaagt cagacccata tgcacttgtg cgtttgggta cccagacatt
     1801 ctgcagtcgt gtcattgatg aagaactcaa cccacagtgg ggagagactt atgaggtgat
     1861 ggtacacgag gtcccagggc aggagattga agtggaggtg ttcgacaagg atccagataa
     1921 agatgacttt ctgggcagaa tgaagctgga tgtagggaag gtgttacagg ctagcgttct
     1981 ggatgattgg ttccctctac aaggtgggca aggccaagtt cacttgaggc tagaatggct
     2041 gtcacttttg tcagatgcag agaaactgga gcaggttcta cagtggaatt ggggagtctc
     2101 ctctcgacca gatcccccgt cagctgccat cttagttgtc tacctggatc gggcccagga
     2161 tcttcctctg aagaagggga acaaggaacc caaccctatg gtacaactgt caattcagga
     2221 tgtgactcag gagagcaagg ctgtctacag taccaactgc ccagtgtggg aggaagcgtt
     2281 ccggttcttc ctacaagacc ctcaaagcca ggagctcgat gtgcaagtga aggatgattc
     2341 cagggccctg actttaggag cactgacgct gcctctggcc cgcctgctga ctgccccaga
     2401 actcatcctg gaccagtggt tccagctcag cagctctggt ccaaactcca gactctatat
     2461 gaaactagtc atgaggatcc tgtacttgga ttcatcagaa atatgcttcc ccacggtgcc
     2521 tggttgtcct ggtgcttggg acgtggacag tgagaatccc cagagaggca gcagtgtgga
     2581 tgccccacct cgaccctgtc acacgactcc tgatagccag tttgggactg agcatgtgct
     2641 tcggatccat gtattagagg cccaggacct gattgccaaa gaccgtttct tggggggact
     2701 ggtgaagggc aagtcagacc cctatgtcaa actaaagttg gcaggacgaa gcttccggag
     2761 ccatgttgtt cgggaagatc tcaatccccg ctggaatgag gtttttgagg tgatcgtcac
     2821 atcagttcca ggccaagagc tagaggttga agtctttgac aaggacttgg acaaggatga
     2881 ttttctgggc aggtgtaaag tgcgtctcac cacagtctta aacagtggct tccttgatga
     2941 gtggctgacc ctggaggatg tcccatctgg ccgcctgcac ttgcgcctgg agcgtctcac
     3001 cccccgtccc actgctgctg agttagagga ggtgctgcag gtgaatagtt tgatccagac
     3061 tcagaagagt gcggagctgg ctgcggccct gctatccatc tatatggagc gggcagagga
     3121 cctcccgctg cgaaaaggca ccaagcacct cagcccttat gctactctca ctgtgggaga
     3181 tagttctcat aaaaccaaga ctatttcgca aacttcagcc cctgtctggg atgagagtgc
     3241 ctcctttctc atcaggaaac cacacactga gagcctagag ttgcaggttc ggggtgaggg
     3301 cactggcgtg ctgggctcat tatccctgcc cctctcagag ctcctcgtgg ctgaccagct
     3361 ctgcttggac cgctggttta cactcagcag tggtcagggg caggtgctac tgagagcaca
     3421 gctagggatc ctggtgtccc agcactcggg agtggaagct catagccaca gctacagcca
     3481 cagctcctca tcgctgagtg aagaaccaga gctctcgggg ggaccccctc acatcacctc
     3541 ctcagcccca gagctccggc agcgcctaac acatgttgac agtccccttg aggctccagc
     3601 cgggcctctg ggccaggtga aactgactct gtggtactac agtgaagaac gaaagctggt
     3661 cagcattgtt catggttgcc ggtcccttcg acagaatgga cgtgatcctc ctgatcccta
     3721 tgtgtcactg ttgctactgc cagacaagaa ccgaggcacc aagaggagga cctcacagaa
     3781 gaagaggacc ctgagtcctg aatttaatga acggtttgag tgggaactcc ccctggatga
     3841 ggcccagaga cgaaagctgg atgtctctgt caagtctaat tcctccttca tgtcaagaga
     3901 gcgtgagctg ctggggaagg tgcagctgga cctagctgag acagaccttt cccagggtgt
     3961 agcccggtgg tatgacctga tggacaacaa ggacaagggc agctcctagc cgcgggcccg
     4021 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     4081 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     4141 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4201 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4261 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     4321 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     4381 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     4441 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     4501 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4561 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     4621 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4681 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     4741 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4801 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4861 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4921 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4981 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5041 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5101 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5161 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5221 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     5281 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5341 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5401 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     5461 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5521 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5581 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5641 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5701 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5761 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     5821 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5881 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5941 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6001 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     6061 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     6121 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6181 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     6241 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     6301 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     6361 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     6421 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     6481 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     6541 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6601 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6661 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6721 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6781 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6841 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6901 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6961 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7021 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7081 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7141 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7201 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7261 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7321 ttatcccctg attctgtgga taaccgtatt accgc
//
