LOCUS       sequence-229124-        6646 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 6646)
  AUTHORS   Fang S, Ferrone M, Yang C, Jensen JP, Tiwari S, Weissman AM
  TITLE     The tumor autocrine motility factor receptor, gp78, is a ubiquitin 
            protein ligase implicated in degradation from the endoplasmic 
            reticulum.
  JOURNAL   Proc Natl Acad Sci U S A. 2001 Dec 4. 98(25):14422-7.
  PUBMED    11724934
REFERENCE   2  (bases 1 to 6646)
  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. 2001 Dec 4. 98(25):14422-7."
COMMENT     Sequence Label: pEGFP-N1 gp78fl
FEATURES             Location/Qualifiers
     source          1..6646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(2583..2585,2586..2588,2589..3302)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2628..2649)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2889..2908)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3236..3257
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3425..3546
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3462..3481)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3516..3535
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3553..4008)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3690..3711)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3902..3921
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4035..4139
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4141..4498
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4141..4332
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4164..4184)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4349..4484
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4533..5327
                     /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             4533..5327
                     /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(4587..4606)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5197..5216
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5515..5534)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5559..5606
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5935..6523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6424..6443
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca
      601 gatccaccat ggtgatgccg ctgctcttcc tcgagcgctt cccctggccc agcctccgca
      661 cctacacggg cctcagcggc ctggccctgc tgggcaccat catcagcgcc taccgcgcgc
      721 tcagccagcc cgaggccggc cccggcgagc cggaccagct aacggcctcg ctgcagcctg
      781 agccgccggc gcccgcccgg ccgagcgccg ggggaccccg ggcccgcgat gtggcccagt
      841 acctgctctc agacagcctc ttcgtgtggg ttctagtaaa taccgcttgc tgtgttttga
      901 tgttggtggc taagctcatc cagtgtattg tgtttggccc tcttcgagtg agtgagagac
      961 agcatctcaa agacaaattt tggaatttta ttttctacaa gttcattttc atctttggtg
     1021 tgctgaatgt ccagacagtg gaggaggtgg tcatgtggtg cctctggttt gccggacttg
     1081 tctttctgca cctgatggtt cagctctgca aggatcgatt tgaatatctt tccttctcgc
     1141 ccaccacgct gatgagcagc cacggtcgag tcctgtccct gttggttgcc atgctgcttt
     1201 cctgctgtgg actggcggcc gtctgctcca tcaccggcta cacccacgga atgcacacct
     1261 tggctttcat ggctgcagag tctcttcttg tgacagtgag gactgctcat gtgattttac
     1321 gatacgtaat tcacctctgg gacctcaacc acgaagggac gtgggaagga aaggggacgt
     1381 atgtctatta cacagactta gtcatggagc tcactctcct gtccctggac ctcatgcacc
     1441 atattcacat gttgttattt ggcaacatct ggttatccat ggccagcctg gtcatcttta
     1501 tgcagctgcg ttacctgttt catgaggtgc aacgtcgaat tcgtcggcac aagaactatc
     1561 tacgtgtggt tggaaacatg gaggccaggt ttgcagttgc aactccagag gagctggctg
     1621 tcaacaatga cgactgtgcc atctgttggg actccatgca ggctgcgcgg aaactgccct
     1681 gtggacatct tttccacaac tcctgtcttc gttcctggct agaacaagac acctcctgtc
     1741 caacatgcag aatgtctctt aatattgccg acaataatcg tgtcagggaa gaacatcaag
     1801 gagagaactt ggatgagaat ttggttcctg tagcagcagc cgaagggaga cctcgcttaa
     1861 accaacacaa tcacttcttc catttcgatg ggtctcggat tgcgagctgg ctgccgagtt
     1921 tttcggttga agtgatgcac accaccaaca ttcttggcat tacgcaggcc agcaactccc
     1981 agctcaatgc aatggctcat cagattcaag agatgtttcc ccaggttcca taccatctgg
     2041 tactgcagga cctccagctg acacgctcag ttgaaataac aacagacaat attttagaag
     2101 gacggattca agtacctttt cctacacagc ggtcagatag catcagacct gcattgaaca
     2161 gtcctgtgga aaggccaagc agtgaccagg aagagggaga aacttctgct cagaccgagc
     2221 gtgtgccact ggacctcagt cctcgcctgg aggagacgct ggacttcggc gaggtggaag
     2281 tggagcccag tgaggtggaa gacttcgagg ctcgtgggag ccgcttctcc aagtctgctg
     2341 atgagagaca gcgcatgctg gtgcagcgta aggacgaact cctccagcaa gctcgcaaac
     2401 gtttcttgaa caaaagttct gaagatgatg cggcctcaga gagcttcctc ccctcggaag
     2461 gtgcgtcctc tgaccccgtg accctgcgtc gaaggatgct ggctgccgcc gcggaacgga
     2521 ggcttcagaa gcagcagacc tcgtcgacgg taccgcgggc ccgggatcca ccggtcgccc
     2581 atatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2641 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2701 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2761 ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga
     2821 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2881 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2941 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3001 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     3061 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     3121 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3181 actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     3241 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     3301 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3361 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3421 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3481 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3541 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3601 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3661 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3721 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3781 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3841 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3901 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3961 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4021 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4081 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4141 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4201 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4261 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4321 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4381 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4441 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4501 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4561 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4621 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4681 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4741 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4801 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4861 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4921 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4981 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5041 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5101 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5161 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5221 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5281 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5341 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5401 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5461 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5521 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5581 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5641 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5701 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5761 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5821 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5881 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5941 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6001 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6061 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6121 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6181 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6241 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6301 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6361 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6421 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6481 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6541 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6601 ccctgattct gtggataacc gtattaccgc catgcattag ttatta
//
