LOCUS       sequence-455035-        7498 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 7498)
  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 7498)
  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: pCI-Neo-gp78/JM20
FEATURES             Location/Qualifiers
     source          1..7498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(49..843)
                     /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(160..179)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     770..789
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(894..1251)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      908..1043
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1060..1251
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1208..1228
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1368..1823)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1505..1526)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1717..1736
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2009..2130
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2046..2065)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2100..2119
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(4247..4265)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4308..4327)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          4310..4442
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(4582..4606)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(4603..4814)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4632..4652)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4815..5194
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5405..5993)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5485..5504)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6164..6955,6956..7024))
                     /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     6787..6806
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7025..7129)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7197..7215
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7253..7275)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7375..7394
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     polyA_signal    7435..7483
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc
       61 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
      121 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
      181 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
      241 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
      301 gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
      361 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
      421 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
      481 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
      541 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
      601 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
      661 ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
      721 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
      781 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
      841 catggtggct ctagccttaa gttcgagact gttgtgtcag aagaatcaag ctttttgcaa
      901 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg
      961 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     1021 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
     1081 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
     1141 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     1201 tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca gaaggtacct
     1261 aaccaagttc ctctttcaga ggttatttca ggccatggtg ctgcgcagat ccgcgtatgc
     1321 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt
     1381 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     1441 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     1501 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     1561 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     1621 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     1681 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     1741 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     1801 taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg
     1861 cgatcggtgc gggcctcttc gctattacgc cagcccggat cgatccttat cggattttac
     1921 cacatttgta gaggttttac ttgctttaaa aaacctccca catctccccc tgaacctgaa
     1981 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2041 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2101 tggtttgtcc aaactcatca atgtatctta tcatgtctgc tcgaagcatt aaccctcact
     2161 aaagggaagc ggccgcccgg gtcgactcta gtaacggccg ccagtgtgct ggaattcggc
     2221 ttcgctagga ggtctgctgc ttctgaagcc tccgttccgc ggcggcagcc agcatccttc
     2281 gacgcagggt cacggggtca gaggacgcac cttccgaggg gaggaagctc tctgaggccg
     2341 catcatcttc agaacttttg ttcaagaaac gtttgcgagc ttgctggagg agttcgtcct
     2401 tacgctgcac cagcatgcgc tgtctctcat cagcagactt ggagaagcgg ctcccacgag
     2461 cctcgaagtc ttccacctca ctgggctcca cttccacctc gccgaagtcc agcgtctcct
     2521 ccaggcgagg actgaggtcc agtggcacac gctcggtctg agcagaagtt tctccctctt
     2581 cctggtcact gcttggcctt tccacaggac tgttcaatgc aggtctgatg ctatctgacc
     2641 gctgtgtagg aaaaggtact tgaatccgtc cttctaaaat attgtctgtt gttatttcaa
     2701 ctgagcgtgt cagctgaagg tcctgcagta ccagatggta tgggacctgg ggaaacatct
     2761 cttgaatctg atgagccatt gcattgagct gggagttgct ggcctgcgta atgccaagaa
     2821 tgttggtggt gtgcatcact tcaaccgaaa aactcggcag ccagctcgca atccgagacc
     2881 catcgaaatg gaagaagtga ttgtgttggt ttaagcgagg tctcccttcg gctgctgcta
     2941 caggaaccaa attctcatcc aagttctctc cttgatgttc ttccctgaca cgattattgt
     3001 cggcaatatt aagagacatt ctgcatgttg gacaggaggt gtcttgttct agccaggaac
     3061 gaagacagga gttgtggaaa agatgtccac agggcagttt ccgcgcagcc tgcatggagc
     3121 cccaacagat ggcacagtcg tcattgttga cagccagctc ctctggagtt gcaactgcaa
     3181 acctggcctc catgtttcca accacacgta gacagttctt gtgccgacga attcgacgtt
     3241 gcacctcatg aaacaggtaa cgcagctgca taaagatgac caggctggcc atggataacc
     3301 agatgttgcc aaataacaac atgtgaatat ggtgcatgag gtccagggac aggagagtga
     3361 gctccatgac taagtctgtg taatagacat acgtcccctt tccttcccac gtcccttcgt
     3421 ggttgaggtc ccagaggtga attacgtatc gtaaaatcac atgagcagtc ctcactgtca
     3481 caagaagaga ctctgcagcc atgaaagcca aggtgtgcat tccgtgggtg tagccggtga
     3541 tggagcagac ggccgccagt ccacagcagg aaagcagcat ggcaaccaac agggacagga
     3601 ctcgaccgtg gctgctcatc agcgtggtgg gcgagaagga aagatattca aatcgatcct
     3661 tgcagagctg aaccatcagg tgcagaaaga caagtccggc aaaccagagg caccacatga
     3721 ccacctcctc cactgtctgg acattcagca caccaaagat gaaaatgaac ttgtagaaaa
     3781 taaaattcca aaatttgtct ttgagatgct gtctctcact cactcgaaga gggccaaaca
     3841 caatacactg gatgagctta gccaccaaca tcaaaacaca gcaagcggta tttactagaa
     3901 cccacacgaa gaggctgtct gagagcaggt actgggccac atcgcgggcc cggggtcccc
     3961 cggcgctcgg ccgggcgggc gccggcggct caggctgcag cgaggccgtt agctggtccg
     4021 gctcgccggg gccggcctcg ggctggctga gcgcgcggta ggcgctgatg atggtgccca
     4081 gcagggccag gccgctgagg cccgtgtagg tgcggaggct gggccagggg aagcgctcga
     4141 ggaagagcag cggcatggcg gcaagccgaa ttctgcagat atccatcaca ctggcggccg
     4201 ctcgagcatg catctagagg taccacgcgt gaattctcga ggctagccta tagtgagtcg
     4261 tattaagtac tctagcctta agagctgtaa ttgaactggg agtggacacc tgtggagaga
     4321 aaggcaaagt ggatgtcagt aagaccaata ggtgcctatc agaaacgcaa gagtcttctc
     4381 tgtctcgaca agcccagttt ctattggtct ccttaaacct gtcttgtaac cttgatactt
     4441 acctgcccag tgcctcacga ccaacttctg caaggctacc ttaagagagt cactgcagct
     4501 taagttcgag actgttgtgt cagaagcact gactgcgtta gcaatttaac tgtgataaac
     4561 taccgcaata aagcttctag tgatctgacg gttcactaaa cgagctctgc ttatatagac
     4621 ctcccaccgt acacgcctac cgcccatttg cgtcaacggg gcgggcgatc gcagttgtta
     4681 cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg
     4741 gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt ggtgtactgc
     4801 caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     4861 aagtcccgta aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     4921 tcaatagggg gcggacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     4981 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     5041 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     5101 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     5161 gattactatt aataactagt caataatcaa tgccaacatg gcggtcatat tggacatgag
     5221 ccaatataaa tgtacatatt atgatataga tacaacgtat gcaatggcca atagccaata
     5281 ttgatttatg ctatataacc aatgaataat atggctaatg gccaatattg aagatctgtc
     5341 gagccatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5401 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5461 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5521 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5581 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5641 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5701 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5761 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5821 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5881 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5941 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6001 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6061 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6121 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6181 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6241 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6301 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6361 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6421 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6481 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6541 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6601 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6661 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6721 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6781 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6841 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6901 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6961 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7021 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7081 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7141 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     7201 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     7261 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     7321 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     7381 cagagcagat tgtactgaga gtgcaccata cgcggatcct tatcgctatc gattcacaca
     7441 aaaaaccaac acacagatgt aatgaaaata aagatatttt attgcggcca tcgtgatg
//
