LOCUS       sequence-207873-        5239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ER marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5239)
  AUTHORS   Zewe JP, Wills RC, Sangappa S, Goulden BD, Hammond GR
  TITLE     SAC1 degrades its lipid substrate PtdIns4P in the endoplasmic 
            reticulum to maintain a steep chemical gradient with donor 
            membranes.
  JOURNAL   Elife. 2018 Feb 20;7. pii: 35588. doi: 10.7554/eLife.35588.
  PUBMED    29461204
REFERENCE   2  (bases 1 to 5239)
  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: "Elife."; 
            date: "2018-02-20"; volume: "7. pii: 35588. doi"; pages: " 
            10.7554/eLife.35588"
COMMENT     Sequence Label: iRFP-Sec61beta
FEATURES             Location/Qualifiers
     source          1..5239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        55..358
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        359..562
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     513..533
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      601..610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             607..1554
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     CDS             1570..1860
                     /codon_start=1
                     /gene="SEC61B"
                     /product="human Sec61-beta subunit of the translocon in the
                     endoplasmic reticulum"
                     /label=Sec61-beta
                     /translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG
                     RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
     polyA_signal    2021..2142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2058..2077)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2112..2131
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2149..2604)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2286..2307)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2498..2517
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2631..2735
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2737..3094
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2737..2928
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2760..2780)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2945..3080
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3129..3923
                     /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             3129..3923
                     /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(3183..3202)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3793..3812
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4111..4130)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4155..4202
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4531..5119
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5020..5039
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
       61 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      121 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      181 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      241 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      301 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      361 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      421 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      481 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      541 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc
      601 gccaccatgg cggaaggatc cgtcgccagg cagcctgacc tcttgacctg cgacgatgag
      661 ccgatccata tccccggtgc catccaaccg catggactgc tgctcgccct cgccgccgac
      721 atgacgatcg ttgccggcag cgacaacctt cccgaactca ccggactggc gatcggcgcc
      781 ctgatcggcc gctctgcggc cgatgtcttc gactcggaga cgcacaaccg tctgacgatc
      841 gccttggccg agcccggggc ggccgtcgga gcaccgatca ctgtcggctt cacgatgcga
      901 aaggacgcag gcttcatcgg ctcctggcat cgccatgatc agctcatctt cctcgagctc
      961 gagcctcccc agcgggacgt cgccgagccg caggcgttct tccgccgcac caacagcgcc
     1021 atccgccgcc tgcaggccgc cgaaaccttg gaaagcgcct gcgccgccgc ggcgcaagag
     1081 gtgcggaaga ttaccggctt cgatcgggtg atgatctatc gcttcgcctc cgacttcagc
     1141 ggcgaagtga tcgcagagga tcggtgcgcc gaggtcgagt caaaactagg cctgcactat
     1201 cctgcctcaa ccgtgccggc gcaggcccgt cggctctata ccatcaaccc ggtacggatc
     1261 attcccgata tcaattatcg gccggtgccg gtcaccccag acctcaatcc ggtcaccggg
     1321 cggccgattg atcttagctt cgccatcctg cgcagcgtct cgcccgtcca tctggaattc
     1381 atgcgcaaca taggcatgca cggcacgatg tcgatctcga ttttgcgcgg cgagcgactg
     1441 tggggattga tcgtttgcca tcaccgaacg ccgtactacg tcgatctcga tggccgccaa
     1501 gcctgcgagc tagtcgccca ggttctggcc tggcagatcg gcgtgatgga agagtccgga
     1561 ctcagatcta tgcctggtcc gacccccagt ggcactaacg tgggatcctc agggcgctct
     1621 cccagcaaag cagtggccgc ccgggcggcg ggatccactg tccggcagag gaaaaatgcc
     1681 agctgtggga caaggagtgc aggccgcaca acctcggcag gcaccggggg gatgtggcga
     1741 ttctacacag aagattcacc tgggctcaaa gttggccctg ttccagtatt ggttatgagt
     1801 cttctgttca tcgcttctgt atttatgttg cacatttggg gcaagtacac tcgttcgtag
     1861 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat
     1921 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     1981 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2041 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2101 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     2161 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2221 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2281 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2341 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2401 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2461 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2521 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2581 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2641 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2701 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     2761 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2821 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2881 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2941 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3001 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3061 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3121 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3181 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3241 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3301 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3361 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3421 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3481 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3541 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3601 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     3661 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3721 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3781 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3841 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3901 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     3961 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4021 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4081 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4141 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     4201 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4261 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4321 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     4381 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4441 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4501 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4561 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4621 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4681 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4741 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4801 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4861 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4921 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4981 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5041 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5101 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5161 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5221 taccgccatg cattagtta
//
