LOCUS       sequence-384518-        5957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Image recycling cargo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5957)
  AUTHORS   Striepen JF, Voeltz GK
  TITLE     Coronin 1C restricts endosomal branched actin to organize ER contact
            and endosome fission.
  JOURNAL   J Cell Biol. 2022 Aug 1;221(8). pii: 213342. doi: 
            10.1083/jcb.202110089. Epub 2022 Jul 8.
  PUBMED    35802042
REFERENCE   2  (bases 1 to 5957)
  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 
            Biol."; date: "2022-08-1"; volume: "221(8). pii: 213342. doi"; 
            pages: " 10.1083/jcb.202110089. Epub 2022 Jul 8"
COMMENT     Sequence Label: FLAG-ADRB2-mNG
FEATURES             Location/Qualifiers
     source          1..5957
                     /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"
     CDS             635..658
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1907..1916
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1913..2623
                     /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      2168..2177
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2745..2866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2782..2801)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2836..2855
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2873..3328)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3010..3031)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3222..3241
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3355..3459
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3461..3818
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3461..3652
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3484..3504)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3669..3804
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3853..4647
                     /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             3853..4647
                     /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(3907..3926)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4517..4536
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4835..4854)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4879..4926
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5255..5843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5744..5763
                     /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 gctagcgcta
      601 ccggactcag atctcgagct caagcttcga aatggactac aaggacgacg atgacaaggg
      661 gcaacccggg aacggcagcg ccttcttgct ggcacccaat agaagccatg cgccggacca
      721 cgacgtcacg cagcaaaggg acgaggtgtg ggtggtgggc atgggcatcg tcatgtctct
      781 catcgtcctg gccatcgtgt ttggcaatgt gctggtcatc acagccattg ccaagttcga
      841 gcgtctgcag acggtcacca actacttcat cacttcactg gcctgtgctg atctggtcat
      901 gggcctggca gtggtgccct ttggggccgc ccatattctt atgaaaatgt ggacttttgg
      961 caacttctgg tgcgagtttt ggacttccat tgatgtgctg tgcgtcacgg ccagcattga
     1021 gaccctgtgc gtgatcgcag tggatcgcta ctttgccatt acttcacctt tcaagtacca
     1081 gagcctgctg accaagaata aggcccgggt gatcattctg atggtgtgga ttgtgtcagg
     1141 ccttacctcc ttcttgccca ttcagatgca ctggtaccgg gccacccacc aggaagccat
     1201 caactgctat gccaatgaga cctgctgtga cttcttcacg aaccaagcct atgccattgc
     1261 ctcttccatc gtgtccttct acgttcccct ggtgatcatg gtcttcgtct actccagggt
     1321 ctttcaggag gccaaaaggc agctccagaa gattgacaaa tctgagggcc gcttccatgt
     1381 ccagaacctt agccaggtgg agcaggatgg gcggacgggg catggactcc gcagatcttc
     1441 caagttctgc ttgaaggagc acaaagccct caagacgtta ggcatcatca tgggcacttt
     1501 caccctctgc tggctgccct tcttcatcgt taacattgtg catgtgatcc aggataacct
     1561 catccgtaag gaagtttaca tcctcctaaa ttggataggc tatgtcaatt ctggtttcaa
     1621 tccccttatc tactgccgga gcccagattt caggattgcc ttccaggagc ttctgtgcct
     1681 gcgcaggtct tctttgaagg cctatgggaa tggctactcc agcaacggca acacagggga
     1741 gcagagtgga tatcacgtgg aacaggagaa agaaaataaa ctgctgtgtg aagacctccc
     1801 aggcacggaa gactttgtgg gccatcaagg tactgtgcct agcgataaca ttgattcaca
     1861 agggaggaat tgtagtacaa atgactcact gctggatcca ccggtcgcca ccatggtgag
     1921 caagggcgag gaggataaca tggcctctct cccagcgaca catgagttac acatctttgg
     1981 ctccatcaac ggtgtggact ttgacatggt gggtcagggc accggcaatc caaatgatgg
     2041 ttatgaggag ttaaacctga agtccaccaa gggtgacctc cagttctccc cctggattct
     2101 ggtccctcat atcgggtatg gcttccatca gtacctgccc taccctgacg ggatgtcgcc
     2161 tttccaggcc gccatggtag atggctccgg ctaccaagtc catcgcacaa tgcagtttga
     2221 agatggtgcc tcccttactg ttaactaccg ctacacctac gagggaagcc acatcaaagg
     2281 agaggcccag gtgaagggga ctggtttccc tgctgacggt cctgtgatga ccaactcgct
     2341 gaccgctgcg gactggtgca ggtcgaagaa gacttacccc aacgacaaaa ccatcatcag
     2401 tacctttaag tggagttaca ccactggaaa tggcaagcgc taccggagca ctgcgcggac
     2461 cacctacacc tttgccaagc caatggcggc taactatctg aagaaccagc cgatgtacgt
     2521 gttccgtaag acggagctca agcactccaa gaccgagctc aacttcaagg agtggcaaaa
     2581 ggcctttacc gatgtgatgg gcatggacga gctgtacaag taagcggccg cgactctaga
     2641 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2701 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2761 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2821 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     2881 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     2941 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3001 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3061 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3121 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3181 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3241 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3301 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3361 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3421 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     3481 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3541 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3601 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3661 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3721 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3781 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     3841 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3901 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3961 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4021 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4081 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4141 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4201 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4261 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4321 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4381 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4441 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4501 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4561 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4621 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4681 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4741 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4801 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     4861 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     4921 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     4981 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5041 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5101 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5161 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5221 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5281 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5341 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5401 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5461 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5521 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5581 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5641 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5701 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5761 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5821 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5881 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5941 gtattaccgc catgcat
//
