LOCUS       sequence-47379-e        7044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7044)
  AUTHORS   Navaroli DM, Bellve KD, Standley C, Lifshitz LM, Cardia J, Lambright
            D, Leonard D, Fogarty KE, Corvera S
  TITLE     Rabenosyn-5 defines the fate of the transferrin receptor following 
            clathrin-mediated endocytosis.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Feb 21;109(8):E471-80. Epub 2012 Jan 
            30.
  PUBMED    22308388
REFERENCE   2  (bases 1 to 7044)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2012-02-21"; volume: "109(8)"; pages: 
            "E471-80. Epub 2012 Jan 30"
COMMENT     Sequence Label: GFP-Rabenosyn-5
FEATURES             Location/Qualifiers
     source          1..7044
                     /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"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3832..3953
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3869..3888)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3923..3942
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3960..4415)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4097..4118)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4309..4328
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4442..4546
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4548..4905
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4548..4739
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4571..4591)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4756..4891
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4940..5734
                     /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             4940..5734
                     /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(4994..5013)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5604..5623
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5922..5941)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5966..6013
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6342..6930
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6831..6850
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcca tggcttctct ggacgaccca
     1381 ggggaagtga gggagggctt cctctgccct ctgtgcctga aggatctgca gtctttctat
     1441 cagcttcact cacattacga ggaagaacac tcaggggaag accgtgatgt caaagggcaa
     1501 attaaaagtc ttgtccagaa ggctaaaaaa gcaaaggaca ggttgttgaa acgagaaggg
     1561 gatgatcgag cagagtcagg gacccaagga tatgagtctt tcagctatgg aggggttgat
     1621 ccttacatgt gggaacccca ggagcttggt gctgtgagaa gccatctttc cgacttcaaa
     1681 aaacaccgag ctgctagaat tgaccactat gttgtggaag tcaataaact aataatcagg
     1741 ttagagaagc tcactgcatt tgacagaaca aatactgagt ctgcaaagat tcgagcaata
     1801 gaaaagtctg tggtgccttg ggtcaacgac caggatgtcc ctttctgtcc agactgtggg
     1861 aataagttca gcatccggaa ccgccgccac cactgccgcc tctgcgggtc tattatgtgc
     1921 aagaagtgta tggagctcat cagccttccc ttggcaaaca agctcaccag tgccagcaag
     1981 gagtccctga gcacccacac cagccccagc cagtcaccca acagtgtcca tggctcccgc
     2041 cgaggcagca tcagcagcat gagcagtgtc agctcggtcc tggatgagaa ggacgatgac
     2101 cggatccgct gctgtacaca ctgcaaggac acgctgctca agagagagca gcagattgat
     2161 gagaaggagc acacacctga catcgtgaag ctctacgaga aattacgact ttgcatggag
     2221 aaagttgacc agaaagctcc agaatacatc aggatggcag catcattaaa tgctggggag
     2281 acaacctaca gtctggaaca tgccagtgac cttcgagtgg aagtgcagaa agtgtatgag
     2341 ctgatagacg ctttaagtaa gaagatctta accttgggct tgaaccagga ccctccacca
     2401 catccaagca atttgcggct gcagagaatg atcagatact cagctacact ttttgtgcag
     2461 gaaaagttgc ttggtttgat gtcactgcca accaaagaac agtttgagga actgaaaaag
     2521 aaaaggaagg aggaaatgga gaggaagagg gccgtggaga gacaagctgc cctggagtcc
     2581 cagcgaaggc ttgaggaaag gcagagtggc ctggcttctc gagcggccaa cggggaggtg
     2641 gcatctctcc gcaggggccc tgcccccttg agaaaggctg agggctggct cccactgtca
     2701 ggaggtcagg ggcagagtga ggactcagac ccgctcctcc agcagatcca caacatcaca
     2761 tcattcatca ggcaggccaa ggccgcgggc cgcatggatg aagtgcgcac tctgcaggag
     2821 aacctgcggc agctgcagga cgagtatgac cagcagcaga cagagaaggc catcgagctg
     2881 tcccggaggc aggctgagga ggaggacctg cagcgggaac agctgcagat gttgcgtgaa
     2941 cgggagttgg aacgagaaag ggagcagttt cgggtggcat ccctgcacac acggactcgg
     3001 tccctggact tcagagaaat cggccctttt cagctggagc ccagcagaga gcctcgcacc
     3061 caccttgctt atgctttgga tctaggctct tccccagttc caagcagcac agctcccaag
     3121 accccttcac ttagctcaac tcaacccacc agagtgtggt ctgggccccc agccgttggc
     3181 caggagcgct taccccagag cagcatgcca cagcaacatg aggggccctc cttaaacccc
     3241 tttgatgagg aagacctctc cagccccatg gaagaggcca ctactggtcc tcctgctgca
     3301 ggggtttcct tagacccttc agcccgcatc ctgaaagagt acaatccttt cgaggaagag
     3361 gacgaggagg aggaagcagt ggcagggaat ccattcattc agccagacag cccagctcct
     3421 aaccccttca gtgaggaaga cgaacatccc cagcagaggc tctcaagccc tctggttcct
     3481 ggtaacccct ttgaggaacc cacctgtatc aacccctttg agatggacag tgacagtggg
     3541 ccagaggctg aggagcccat agaggaagag ctcctcctgc agcagatcga taacatcaag
     3601 gcatacatct ttgatgccaa gcagtgcggc cgcctggatg aggtagaggt gctgacagag
     3661 aatctgcggg agctgaagca caccctggcc aagcagaagg ggggcactga ctgatctaga
     3721 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3781 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3841 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3901 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     3961 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     4021 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     4081 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     4141 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     4201 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     4261 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     4321 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     4381 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     4441 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4501 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     4561 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4621 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4681 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4741 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4801 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4861 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     4921 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4981 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     5041 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     5101 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5161 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5221 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5281 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5341 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5401 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5461 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5521 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5581 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5641 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5701 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5761 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5821 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5881 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     5941 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     6001 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     6061 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     6121 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     6181 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6241 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6301 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6361 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6421 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6481 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6541 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6601 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6661 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6721 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6781 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6841 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6901 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6961 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7021 gataaccgta ttaccgccat gcat
//
