LOCUS       sequence-396460-        6028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6028)
  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 6028)
  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: pCINeo CD3 delta
FEATURES             Location/Qualifiers
     source          1..6028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1091..1100
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1622..1648
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1732..1853
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1743..1762)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1797..1816
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2039..2494
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2126..2145)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2336..2357
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2611..2968
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2611..2802
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2634..2654)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2819..2954
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3019..3813
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3073..3092)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3683..3702
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3877..3925
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3966..3985)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4085..4107
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4145..4163)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4231..4335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4336..4404,4405..5196)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4554..4573)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5367..5955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5856..5875
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc gccaccatgg aacacagcgg gattctggct agtctgatac tgattgctgt
     1141 tctcccccaa gggagcccct tcaagataca agtgaccgaa tatgaggaca aagtatttgt
     1201 gacctgcaat accagcgtca tgcatctaga tggaacggtg gaaggatggt ttgcaaagaa
     1261 taaaacactc aacttgggca aaggcgttct ggacccacga gggatatatc tgtgtaatgg
     1321 gacagagcag ctggcaaagg tggtgtcttc tgtgcaagtc cattaccgaa tgtgccagaa
     1381 ctgtgtggag ctagactcgg gcaccatggc tggtgtcatc ttcattgacc tcatcgcaac
     1441 tctgctcctg gctttgggcg tctactgctt tgcaggacat gagaccggaa ggccttctgg
     1501 ggctgctgag gttcaagcac tgctgaagaa tgagcagctg tatcagcctc ttcgagatcg
     1561 tgaagatacc cagtacagcc gtcttggagg gaactggccc cggaacaaga aatctggagg
     1621 ttacccatac gacgtcccag actacgctta agaattcacg cgtggtacct ctagagtcga
     1681 cccgggcggc cgcttccctt tagtgagggt taatgcttcg agcagacatg ataagataca
     1741 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1801 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1861 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca
     1921 agtaaaacct ctacaaatgt ggtaaaatcc gataaggatc gatccgggct ggcgtaatag
     1981 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac
     2041 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2101 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2161 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2221 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2281 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2341 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2401 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2461 gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     2521 atctgtgcgg tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac
     2581 ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt
     2641 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2701 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     2761 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     2821 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     2881 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     2941 cttttttgga ggcctaggct tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa
     3001 cttaaggcta gagccaccat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3061 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3121 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3181 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3241 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3301 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3361 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3421 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3481 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3541 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3601 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3661 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3721 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3781 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     3841 accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta
     3901 catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatccgcgt atggtgcact
     3961 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     4021 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4081 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     4141 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4201 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4261 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4321 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4381 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4441 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4501 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4561 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4621 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4681 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4741 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4801 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4861 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4921 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4981 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5041 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5101 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5161 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5221 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5281 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5341 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5401 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5461 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5521 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5581 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5641 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5701 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5761 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5821 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5881 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5941 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6001 ccttttgctc acatggctcg acagatct
//
