LOCUS       sequence-47378-e        7047 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 7047)
  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 7047)
  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: TagRFP-T-Rabenosyn-5
FEATURES             Location/Qualifiers
     source          1..7047
                     /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             613..1344
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    3835..3956
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3872..3891)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3926..3945
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3963..4418)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4100..4121)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4312..4331
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4445..4549
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4551..4908
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4551..4742
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4574..4594)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4759..4894
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4943..5737
                     /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             4943..5737
                     /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(4997..5016)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5607..5626
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5925..5944)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5969..6016
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6345..6933
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6834..6853
                     /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 ccatggtgtc taagggcgaa gagctgatta aggagaacat gcacatgaag
      661 ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc
      721 aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc
      781 ttcgccttcg acatcctggc taccagcttc atgtacggca gcagaacctt catcaaccac
      841 acccagggca tccccgattt ctttaagcag tccttccctg agggcttcac atgggagaga
      901 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
      961 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg
     1021 atgcagaaga aaacactcgg ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc
     1081 ggcctggaag gcagaaccga catggccctg aagctcgtgg gcgggggcca cctgatctgc
     1141 aacttcaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc
     1201 tactatgtgg accacagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag
     1261 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaactt
     1321 aatggcatgg acgagctgta caagactcga gctcaagctt ccatggcttc tctggacgac
     1381 ccaggggaag tgagggaggg cttcctctgc cctctgtgcc tgaaggatct gcagtctttc
     1441 tatcagcttc actcacatta cgaggaagaa cactcagggg aagaccgtga tgtcaaaggg
     1501 caaattaaaa gtcttgtcca gaaggctaaa aaagcaaagg acaggttgtt gaaacgagaa
     1561 ggggatgatc gagcagagtc agggacccaa ggatatgagt ctttcagcta tggaggggtt
     1621 gatccttaca tgtgggaacc ccaggagctt ggtgctgtga gaagccatct ttccgacttc
     1681 aaaaaacacc gagctgctag aattgaccac tatgttgtgg aagtcaataa actaataatc
     1741 aggttagaga agctcactgc atttgacaga acaaatactg agtctgcaaa gattcgagca
     1801 atagaaaagt ctgtggtgcc ttgggtcaac gaccaggatg tccctttctg tccagactgt
     1861 gggaataagt tcagcatccg gaaccgccgc caccactgcc gcctctgcgg gtctattatg
     1921 tgcaagaagt gtatggagct catcagcctt cccttggcaa acaagctcac cagtgccagc
     1981 aaggagtccc tgagcaccca caccagcccc agccagtcac ccaacagtgt ccatggctcc
     2041 cgccgaggca gcatcagcag catgagcagt gtcagctcgg tcctggatga gaaggacgat
     2101 gaccggatcc gctgctgtac acactgcaag gacacgctgc tcaagagaga gcagcagatt
     2161 gatgagaagg agcacacacc tgacatcgtg aagctctacg agaaattacg actttgcatg
     2221 gagaaagttg accagaaagc tccagaatac atcaggatgg cagcatcatt aaatgctggg
     2281 gagacaacct acagtctgga acatgccagt gaccttcgag tggaagtgca gaaagtgtat
     2341 gagctgatag acgctttaag taagaagatc ttaaccttgg gcttgaacca ggaccctcca
     2401 ccacatccaa gcaatttgcg gctgcagaga atgatcagat actcagctac actttttgtg
     2461 caggaaaagt tgcttggttt gatgtcactg ccaaccaaag aacagtttga ggaactgaaa
     2521 aagaaaagga aggaggaaat ggagaggaag agggccgtgg agagacaagc tgccctggag
     2581 tcccagcgaa ggcttgagga aaggcagagt ggcctggctt ctcgagcggc caacggggag
     2641 gtggcatctc tccgcagggg ccctgccccc ttgagaaagg ctgagggctg gctcccactg
     2701 tcaggaggtc aggggcagag tgaggactca gacccgctcc tccagcagat ccacaacatc
     2761 acatcattca tcaggcaggc caaggccgcg ggccgcatgg atgaagtgcg cactctgcag
     2821 gagaacctgc ggcagctgca ggacgagtat gaccagcagc agacagagaa ggccatcgag
     2881 ctgtcccgga ggcaggctga ggaggaggac ctgcagcggg aacagctgca gatgttgcgt
     2941 gaacgggagt tggaacgaga aagggagcag tttcgggtgg catccctgca cacacggact
     3001 cggtccctgg acttcagaga aatcggccct tttcagctgg agcccagcag agagcctcgc
     3061 acccaccttg cttatgcttt ggatctaggc tcttccccag ttccaagcag cacagctccc
     3121 aagacccctt cacttagctc aactcaaccc accagagtgt ggtctgggcc cccagccgtt
     3181 ggccaggagc gcttacccca gagcagcatg ccacagcaac atgaggggcc ctccttaaac
     3241 ccctttgatg aggaagacct ctccagcccc atggaagagg ccactactgg tcctcctgct
     3301 gcaggggttt ccttagaccc ttcagcccgc atcctgaaag agtacaatcc tttcgaggaa
     3361 gaggacgagg aggaggaagc agtggcaggg aatccattca ttcagccaga cagcccagct
     3421 cctaacccct tcagtgagga agacgaacat ccccagcaga ggctctcaag ccctctggtt
     3481 cctggtaacc cctttgagga acccacctgt atcaacccct ttgagatgga cagtgacagt
     3541 gggccagagg ctgaggagcc catagaggaa gagctcctcc tgcagcagat cgataacatc
     3601 aaggcataca tctttgatgc caagcagtgc ggccgcctgg atgaggtaga ggtgctgaca
     3661 gagaatctgc gggagctgaa gcacaccctg gccaagcaga aggggggcac tgactgatct
     3721 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3781 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3841 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3901 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     3961 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     4021 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     4081 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     4141 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     4201 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     4261 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     4321 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     4381 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     4441 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4501 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     4561 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4621 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4681 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4741 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4801 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4861 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4921 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4981 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5041 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5101 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     5161 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5221 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5281 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5341 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5401 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5461 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5521 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5581 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5641 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5701 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5761 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5821 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5881 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5941 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     6001 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     6061 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     6121 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     6181 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6241 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6301 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6361 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6421 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6481 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6541 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6601 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6661 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6721 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6781 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6841 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6901 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6961 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     7021 gtggataacc gtattaccgc catgcat
//
