LOCUS       sequence-451899-        8361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Sec31A protein labeled with mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8361)
  AUTHORS   Westrate LM, Hoyer MJ, Nash MJ, Voeltz GK
  TITLE     Vesicular and uncoated Rab1-dependent cargo carriers facilitate ER 
            to Golgi transport.
  JOURNAL   J Cell Sci. 2020 Jul 24;133(14). pii: jcs.239814. doi: 
            10.1242/jcs.239814.
  PUBMED    32616562
REFERENCE   2  (bases 1 to 8361)
  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: "J Cell 
            Sci."; date: "2020-07-24"; volume: "133(14). pii: jcs.239814. doi"; 
            pages: " 10.1242/jcs.239814"
COMMENT     Sequence Label: mNeonGreen-Sec31A
FEATURES             Location/Qualifiers
     source          1..8361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..25
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        280..583
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        584..787
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     738..758
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      826..835
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             832..1539
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      1087..1096
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5368..5489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5405..5424)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5459..5478
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5496..5951)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5633..5654)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5845..5864
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5978..6082
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6084..6441
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6084..6275
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6107..6127)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6292..6427
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6476..7270
                     /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             6476..7270
                     /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(6530..6549)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7140..7159
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7458..7477)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7502..7549
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(7878..8361,1..105)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
       61 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
      121 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
      181 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
      241 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      301 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      361 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      421 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      481 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      541 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      601 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      661 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      721 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      781 cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggtcgccac catggtgagc
      841 aagggcgagg aggataacat ggcctctctc ccagcgacac atgagttaca catctttggc
      901 tccatcaacg gtgtggactt tgacatggtg ggtcagggca ccggcaatcc aaatgatggt
      961 tatgaggagt taaacctgaa gtccaccaag ggtgacctcc agttctcccc ctggattctg
     1021 gtccctcata tcgggtatgg cttccatcag tacctgccct accctgacgg gatgtcgcct
     1081 ttccaggccg ccatggtaga tggctccggc taccaagtcc atcgcacaat gcagtttgaa
     1141 gatggtgcct cccttactgt taactaccgc tacacctacg agggaagcca catcaaagga
     1201 gaggcccagg tgaaggggac tggtttccct gctgacggtc ctgtgatgac caactcgctg
     1261 accgctgcgg actggtgcag gtcgaagaag acttacccca acgacaaaac catcatcagt
     1321 acctttaagt ggagttacac cactggaaat ggcaagcgct accggagcac tgcgcggacc
     1381 acctacacct ttgccaagcc aatggcggct aactatctga agaaccagcc gatgtacgtg
     1441 ttccgtaaga cggagctcaa gcactccaag accgagctca acttcaagga gtggcaaaag
     1501 gcctttaccg atgtgatggg catggacgag ctgtacaagt ccggactcag atctcgagct
     1561 caagcttcga attcgatgaa gttaaaggaa gtagatcgta cagccatgca ggcatggagc
     1621 cctgcccaga atcaccccat ttacctagca acaggaacat ctgctcagca attggatgca
     1681 acatttagta cgaatgcttc ccttgagata tttgaattag acctctctga tccatccttg
     1741 gatatgaaat cttgtgccac attctcctct tctcacaggt accacaagtt gatttggggg
     1801 ccttataaaa tggattccaa aggagatgtc tctggagttc tgattgcagg tggtgaaaat
     1861 ggaaatatta ttctctatga tccttctaaa attatagctg gagacaagga agttgtgatt
     1921 gcccagaatg acaagcatac tggcccagtg agagccttgg atgtgaacat tttccagact
     1981 aatctggtag cttctggtgc taatgaatct gaaatctaca tatgggatct aaataatttt
     2041 gcaaccccaa tgacaccagg agccaaaaca cagccgccag aagatatcag ctgcattgca
     2101 tggaacagac aagttcagca tattttagca tcagccagtc ccagtggccg ggccactgta
     2161 tgggatctta ggaaaaatga gccaatcatc aaagtcagtg accatagtaa cagaatgcat
     2221 tgttctgggt tggcatggca tcctgatgtt gctactcaga tggtccttgc ctccgaggat
     2281 gaccggttac cagtgatcca gatgtgggat cttcgatttg cttcctctcc acttcgtgtc
     2341 ctggaaaacc atgccagggg gattttggca attgcttgga gcatggcaga tcctgaattg
     2401 ttactgagct gtggaaaaga tgctaagatt ctctgctcca atccaaacac aggagaggtg
     2461 ttatatgaac ttcccaccaa cacacagtgg tgcttcgata ttcagtggtg tccccgaaat
     2521 cctgctgtct tatcagctgc ttcgtttgat gggcgtatca gtgtttattc tatcatggga
     2581 ggaagcacag atggtttaag acagaaacaa gttgacaagc tttcatcatc ttttgggaat
     2641 cttgatccct ttggcacagg acagcccctt cctccgttac aaattccaca gcagactgct
     2701 cagcatagta tagtgctgcc tctgaagaag ccgcccaagt ggattcgaag gcctgttggt
     2761 gcttcttttt catttggagg caaactggtt acgtttgaga atgtcagaat gccttctcat
     2821 cagggagctg agcagcagca gcagcagcac catgtgttca ttagtcaggt tgtaacagaa
     2881 aaggagttcc tcagccgatc agaccaactt cagcaggctg tgcagtcaca aggatttatc
     2941 aattattgcc aaaaaaaaat tgatgcttct cagactgaat ttgagaaaaa tgtgtggtcc
     3001 tttttgaagg taaactttga ggatgattct cgtggaaaat accttgaact tctaggatac
     3061 agaaaagaag atctaggaaa gaagattgct ttggccttga acaaagtgga tggagccaat
     3121 gtggctctta aagactctga ccaagtagca cagagtgatg gggaggagag ccctgctgct
     3181 gaagagcagc tcttgggaga gcacattaaa gaggaaaaag aagaatctga atttctaccc
     3241 tcatctggag gaacatttaa tatctctgtc agtggggaca ttgatggttt aattactcag
     3301 gctttgctga cgggcaattt tgagagtgct gttgaccttt gtttacatga taaccgcatg
     3361 gccgatgcca ttatattggc catagcaggt ggacaagaac tcttggctcg aacccagaaa
     3421 aaatacttcg caaaatccca aagcaaaatt accaggctca tcactgcagt ggtgatgaag
     3481 aactggaaag agattgttga gtcttgtgat cttaaaaatt ggagagaggc tttagctgca
     3541 gtattgactt atgcaaagcc ggatgaattt tcagcccttt gtgatctttt gggaaccagg
     3601 cttgaaaatg aaggagatag cctcctgcag actcaagcat gtctctgcta tatttgtgcg
     3661 gggaatgtag agaaattagt tgcatgttgg actaaagctc aagatggaag ccaccctttg
     3721 tcacttcagg atctgattga gaaagttgtc atcctgcgaa aagctgtgca actcactcaa
     3781 gccatggaca ctagtactgt aggagttctc ttggctgcga agatgagtca gtatgccaat
     3841 ttgttggcag ctcagggcag tattgctgca gccttggctt ttcttcctga caacaccaac
     3901 cagccaaata tcatgcagct tcgtgacaga ctttgtagag cacaaggaga gcctgtagca
     3961 ggacatgaat cacctaaaat tccgtacgag aaacagcagc tccccaaggg caggcctgga
     4021 ccagttgctg gccaccacca gatgccaaga gttcaaactc aacaatatta tccccatgga
     4081 gaaaatcctc cacctccggg tttcataatg catggaaatg ttaatccaaa tgctgctggt
     4141 cagcttccca catctccagg tcatatgcac acccaggtac caccttatcc acagccacag
     4201 ccttatcaac cagcccagcc gtatcccttc ggaacagggg ggtcagcaat gtatcgacct
     4261 cagcagcctg ttgctcctcc tacttcaaac gcttacccta acacccctta catatcttct
     4321 gcttcttcct atactgggca gtctcagctg tacgcagcac agcaccaggc ctcttcacct
     4381 acctccagcc ctgctacttc tttccctcct cccccttcct ctggagcatc cttccagcat
     4441 ggcggaccag gagctccacc atcatcttca gcttatgcac tgcctcctgg aacaacaggt
     4501 acactgcctg ctgccagtga gctgcctgcg tcccaaagaa caggtcctca gaatggttgg
     4561 aatgaccctc cagctttgaa cagagtaccc aaaaagaaga agatgcctga aaacttcatg
     4621 cctcctgttc ccatcacatc accaatcatg aacccgttgg gtgaccccca gtcacaaatg
     4681 ctgcagcaac agccttcagc tccagtacca ctgtcaagcc agtcttcatt cccacagcca
     4741 catcttccag gtggccagcc cttccatggc gtacagcaac ctcttggtca aacaggcatg
     4801 ccaccatctt tttcaaagcc caatattgaa ggtgccccag gggctcctat tggaaatacc
     4861 ttccagcatg tgcagtcttt gccaacaaaa aaaattacca agaaacctat tccagatgag
     4921 cacctcattc taaagaccac atttgaggat cttattcagc gctgcctttc ttcagcaaca
     4981 gaccctcaaa ccaagaggaa gctagatgat gccagcaaac gtttggagtt tctgtatgat
     5041 aaacttaggg aacagacact ttcaccaaca atcaccagtg gtttacacaa cattgcaagg
     5101 agcattgaaa ctcgaaacta ctcagaagga ttgaccatgc atacccacat agttagcacc
     5161 agcaacttca gtgagacctc tgctttcatg ccagttctca aagttgttct cacccaggcc
     5221 aataagctgg gtgtctaagg atccaccgga tctagataac tgatcataat cagccatacc
     5281 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     5341 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     5401 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5461 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     5521 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5581 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     5641 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5701 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     5761 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5821 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     5881 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     5941 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6001 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6061 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     6121 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6181 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     6241 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     6301 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6361 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6421 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     6481 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     6541 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     6601 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     6661 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     6721 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     6781 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     6841 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     6901 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     6961 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     7021 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     7081 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     7141 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     7201 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     7261 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     7321 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     7381 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     7441 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     7501 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     7561 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     7621 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     7681 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     7741 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     7801 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     7861 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     7921 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     7981 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     8041 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     8101 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8161 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     8221 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     8281 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     8341 ggagagcgca cgagggagct t
//
