LOCUS       sequence-494680-        7267 bp    DNA     circular SYN 04-DEC-2025
DEFINITION  Expresses Rtn4b connected with C-terminus AcGFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7267)
  AUTHORS   Wang B, Zhao Z, Xiong M, Yan R, Xu K
  TITLE     The endoplasmic reticulum adopts two distinct tubule forms.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 May 3;119(18):e2117559119. doi: 
            10.1073/pnas.2117559119. Epub 2022 Apr 26.
  PUBMED    35471903
REFERENCE   2  (bases 1 to 7267)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2022-05-3"; volume: "119(18):e2117559119. 
            doi"; pages: " 10.1073/pnas.2117559119. Epub 2022 Apr 26"
COMMENT     Sequence Label: Rtn4b-GFP
FEATURES             Location/Qualifiers
     source          1..7267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2107..2826
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     primer_bind     complement(2858..2875)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2864..3088
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3134..3562
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3221..3240)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3431..3452
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3571..3591)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3576..3905
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3756..3891
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3818..3837
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3972..4766
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4026..4045)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4636..4655
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4940..5061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4977..4996)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5031..5050
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5110..5126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5110..5126)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5123..5145)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5134..5150
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5158..5164,5165..5182,5183..5188))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5203..5224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5341..5358)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5512..6100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5592..5611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6271..7062,7063..7131))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6894..6913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7132..7236)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcatg
      961 gaagacctgg accagtctcc tctggtctcg tcctcggaca gcccaccccg gccgcagccc
     1021 gcgttcaagt accagttcgt gagggagccc gaggacgagg aggaagaaga ggaggaggaa
     1081 gaggaggacg aggacgaaga cctggaggag ctggaggtgc tggagaggaa gcccgccgcc
     1141 gggctgtccg cggccccagt gcccaccgcc cctgccgccg gcgcgcccct gatggacttc
     1201 ggaaatgact tcgtgccgcc ggcgccccgg ggacccctgc cggccgctcc ccccgtcgcc
     1261 ccggagcggc agccgtcttg ggacccgagc ccggtgtcgt cgaccgtgcc cgcgccatcc
     1321 ccgctgtctg ctgccgcagt ctcgccctcc aagctccctg aggacgacga gcctccggcc
     1381 cggcctcccc ctcctccccc ggccagcgtg agcccccagg cagagcccgt gtggaccccg
     1441 ccagccccgg ctcccgccgc gcccccctcc accccggccg cgcccaagcg caggggctcc
     1501 tcgggctcag tggttgttga cctcctgtac tggagagaca ttaagaagac tggagtggtg
     1561 tttggtgcca gcctattcct gctgctttca ttgacagtat tcagcattgt gagcgtaaca
     1621 gcctacattg ccttggccct gctctctgtg accatcagct ttaggatata caagggtgtg
     1681 atccaagcta tccagaaatc agatgaaggc cacccattca gggcatatct ggaatctgaa
     1741 gttgctatat ctgaggagtt ggttcagaag tacagtaatt ctgctcttgg tcatgtgaac
     1801 tgcacgataa aggaactcag gcgcctcttc ttagttgatg atttagttga ttctctgaag
     1861 tttgcagtgt tgatgtgggt atttacctat gttggtgcct tgtttaatgg tctgacacta
     1921 ctgattttgg ctctcatttc actcttcagt gttcctgtta tttatgaacg gcatcaggca
     1981 cagatagatc attatctagg acttgcaaat aagaatgtta aagatgctat ggctaaaatc
     2041 caagcaaaaa tccctggatt gaagcgcaaa gctgaaacgg taccgcgggc ccgggatcca
     2101 ccggtcatgg tgagcaaggg cgccgagctg ttcaccggca tcgtgcccat cctgatcgag
     2161 ctgaatggcg atgtgaatgg ccacaagttc agcgtgagcg gcgagggcga gggcgatgcc
     2221 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc tgtgccctgg
     2281 cccaccctgg tgaccaccct gagctacggc gtgcagtgct tctcacgcta ccccgatcac
     2341 atgaagcagc acgacttctt caagagcgcc atgcctgagg gctacatcca ggagcgcacc
     2401 atcttcttcg aggatgacgg caactacaag tcgcgcgccg aggtgaagtt cgagggcgat
     2461 accctggtga atcgcatcga gctgaccggc accgatttca aggaggatgg caacatcctg
     2521 ggcaataaga tggagtacaa ctacaacgcc cacaatgtgt acatcatgac cgacaaggcc
     2581 aagaatggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggatgg cagcgtgcag
     2641 ctggccgacc actaccagca gaataccccc atcggcgatg gccctgtgct gctgcccgat
     2701 aaccactacc tgtccaccca gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac
     2761 atgatctact tcggcttcgt gaccgccgcc gccatcaccc acggcatgga tgagctgtac
     2821 aagtgatcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2881 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2941 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3001 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3061 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3121 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3181 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3241 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3301 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3361 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3421 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3481 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3541 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3601 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3661 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3721 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3781 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3841 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3901 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3961 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4021 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4081 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4141 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4201 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4261 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4321 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4381 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4441 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4501 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4561 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4621 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4681 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4741 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4801 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4861 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4921 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4981 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5041 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5101 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5161 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5221 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5281 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5341 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5401 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5461 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5521 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5581 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5641 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5701 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5761 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5821 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5881 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5941 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6001 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6061 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6121 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6181 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6241 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6301 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6361 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6421 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6481 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6541 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6601 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6661 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6721 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6781 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6841 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6901 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6961 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7021 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7081 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7141 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7201 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7261 tgacgtc
//
