LOCUS       sequence-483845-        5152 bp    DNA     circular SYN 19-AUG-2025
DEFINITION  Mammalian expression of HyPerFLEX targeted to the luminal side of 
            the endoplasmic reticulum membranes .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5152)
  TITLE     Belousov lab plasmids 2025
REFERENCE   2  (bases 1 to 5152)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pC1-HyPerFLEX-ERlum
FEATURES             Location/Qualifiers
     source          1..5152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1940..2061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1977..1996)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2031..2050
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2068..2523)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2205..2226)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2417..2436
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2550..2654
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2656..3013
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2656..2847
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2679..2699)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2864..2999
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3048..3842
                     /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             3048..3842
                     /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(3102..3121)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3712..3731
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4030..4049)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4074..4121
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4450..5038
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4939..4958
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgc
      601 tgctatccgt gccgttgctg ctcggcctcc tcggcctggc cgtcgccgga ccggtcgcca
      661 cccacctggc taatgaggag caaaacgaat tggaaggtgc ttttaagctg ggcttgatct
      721 ttaccgtcgc cccgtacttg ctgcctaagt tgatcgtcag cctgcgccgc accgccccga
      781 agatgccttt gatgttggag gagaattaca cccacaccct taccgagtca ctgaaacgcg
      841 gcgacgttga cgccatcatt gtggcagagc cgtttcaaga gccgggtatt gtgactgaac
      901 ctttgtatga cgagccgttt ttcgttatcg ttccgaaggg ccatagcttc gaggagttgg
      961 atgccgttag ccctcgcatg ttgggcgaag aacaagttct gcttctgacc gagggcaact
     1021 gtatgcgtga ccaggtgctt agcattttta ccaaggtcaa ggtgcacatg aagaaagccc
     1081 tttccggtga cagctattgg gtctttgtga aacgggtagg tggcactggc ggtgagcatg
     1141 ttgtctttgg cagtgaggac atcgagaaca ctatggccaa aatggacgac ggacaactgg
     1201 atgggttggc ctttggcgca attcagctcg atggtgacgg gaatatcctg cagtacaatg
     1261 ctgctgaagg agacatcaca ggcagagatc ccaaacaggt gattgggaag aacttcttca
     1321 aggatgttgc acctggaacg gattctcccg agttttacgg caaattcaag gaaggcgtag
     1381 cgtcagggaa tctgaacacc atgttcgaat ggatgatacc gactcttacc gatcatagtt
     1441 gcagcgagct tgccgccaaa cagcgcattc agggtctcac caacaccctt caaggctcaa
     1501 gcatcgatac catccgccac atggttgcca gtggcttggc aatcagcgtc ttgcctgcca
     1561 ctgcccttac cgaaaacgac cacatgctgt ttagcattat cccgtttgaa ggcaccccgc
     1621 ctagccgccg cgttgtcctg gcctatcgcc gcaactttgt ccgccctaag gccctttcag
     1681 ccatgaaagc cgcaattatg caaagccaac tgcatggtgt tagcttcatc tgcgatggtg
     1741 gaggaggtag atctggaaag gacgagctgt aaaagcttcg aattctgcag tcgacggtac
     1801 cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
     1861 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     1921 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     1981 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2041 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     2101 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     2161 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     2221 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2281 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     2341 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2401 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     2461 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2521 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2581 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2641 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     2701 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2761 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2821 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2881 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2941 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3001 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3061 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3121 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3181 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     3241 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3301 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3361 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3421 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3481 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3541 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     3601 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     3661 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     3721 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     3781 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     3841 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     3901 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     3961 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     4021 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     4081 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4141 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4201 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4261 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     4321 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4381 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4441 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4501 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4561 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4621 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4681 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4741 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4801 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4861 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4921 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4981 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5041 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5101 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
