LOCUS       sequence-420916-        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-gp78/2853
FEATURES             Location/Qualifiers
     source          1..7498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(644..662)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(705..724)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          707..839
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(979..1003)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1000..1211)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1029..1049)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1212..1591
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1802..2390)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1882..1901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2561..3352,3353..3421))
                     /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     3184..3203
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3422..3526)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3594..3612
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3650..3672)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3772..3791
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     polyA_signal    3832..3880
                     /note="synthetic polyadenylation signal"
     CDS             complement(3944..4738)
                     /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(4055..4074)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4665..4684
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4789..5146)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4803..4938
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4955..5146
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5103..5123
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5263..5718)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5400..5421)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5612..5631
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5904..6025
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5941..5960)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5995..6014
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gaccgtggct gctcatcagc gtggtgggcg agaaggaaag atattcaaat cgatccttgc
       61 agagctgaac catcaggtgc agaaagacaa gtccggcaaa ccagaggcac cacatgacca
      121 cctcctccac tgtctggaca ttcagcacac caaagatgaa aatgaacttg tagaaaataa
      181 aattccaaaa tttgtctttg agatgctgtc tctcactcac tcgaagaggg ccaaacacaa
      241 tacactggat gagcttagcc accaacatca aaacacagca agcggtattt actagaaccc
      301 acacgaagag gctgtctgag agcaggtact gggccacatc gcgggcccgg ggtcccccgg
      361 cgctcggccg ggcgggcgcc ggcggctcag gctgcagcga ggccgttagc tggtccggct
      421 cgccggggcc ggcctcgggc tggctgagcg cgcggtaggc gctgatgatg gtgcccagca
      481 gggccaggcc gctgaggccc gtgtaggtgc ggaggctggg ccaggggaag cgctcgagga
      541 agagcagcgg catggcggca agccgaattc tgcagatatc catcacactg gcggccgctc
      601 gagcatgcat ctagaggtac cacgcgtgaa ttctcgaggc tagcctatag tgagtcgtat
      661 taagtactct agccttaaga gctgtaattg aactgggagt ggacacctgt ggagagaaag
      721 gcaaagtgga tgtcagtaag accaataggt gcctatcaga aacgcaagag tcttctctgt
      781 ctcgacaagc ccagtttcta ttggtctcct taaacctgtc ttgtaacctt gatacttacc
      841 tgcccagtgc ctcacgacca acttctgcaa ggctacctta agagagtcac tgcagcttaa
      901 gttcgagact gttgtgtcag aagcactgac tgcgttagca atttaactgt gataaactac
      961 cgcaataaag cttctagtga tctgacggtt cactaaacga gctctgctta tatagacctc
     1021 ccaccgtaca cgcctaccgc ccatttgcgt caacggggcg ggcgatcgca gttgttacga
     1081 cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg
     1141 tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattggt gtactgccaa
     1201 aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag
     1261 tcccgtaagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
     1321 atagggggcg gacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt
     1381 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac
     1441 gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc
     1501 gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat
     1561 tactattaat aactagtcaa taatcaatgc caacatggcg gtcatattgg acatgagcca
     1621 atataaatgt acatattatg atatagatac aacgtatgca atggccaata gccaatattg
     1681 atttatgcta tataaccaat gaataatatg gctaatggcc aatattgaag atctgtcgag
     1741 ccatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1801 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1861 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1921 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1981 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2041 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2101 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2161 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2221 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2281 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2341 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2401 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2461 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2521 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2581 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2641 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2701 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2761 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2821 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2881 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2941 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3001 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3061 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3121 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3181 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3241 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3301 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3361 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3421 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3481 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3541 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     3601 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3661 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     3721 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     3781 agcagattgt actgagagtg caccatacgc ggatccttat cgctatcgat tcacacaaaa
     3841 aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg tgatggcagg
     3901 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     3961 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     4021 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     4081 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     4141 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     4201 cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     4261 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     4321 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     4381 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     4441 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     4501 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca
     4561 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     4621 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     4681 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgg
     4741 tggctctagc cttaagttcg agactgttgt gtcagaagaa tcaagctttt tgcaaaagcc
     4801 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     4861 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     4921 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     4981 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     5041 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     5101 acaccctaac tgacacacat tccacagctg gttctttccg cctcagaagg tacctaacca
     5161 agttcctctt tcagaggtta tttcaggcca tggtgctgcg cagatccgcg tatgcggtgt
     5221 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata
     5281 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     5341 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     5401 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     5461 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     5521 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     5581 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     5641 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     5701 cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5761 ggtgcgggcc tcttcgctat tacgccagcc cggatcgatc cttatcggat tttaccacat
     5821 ttgtagaggt tttacttgct ttaaaaaacc tcccacatct ccccctgaac ctgaaacata
     5881 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     5941 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6001 tgtccaaact catcaatgta tcttatcatg tctgctcgaa gcattaaccc tcactaaagg
     6061 gaagcggccg cccgggtcga ctctagtaac ggccgccagt gtgctggaat tcggcttcgc
     6121 taggaggtct gctgcttctg aagcctccgt tccgcggcgg cagccagcat ccttcgacgc
     6181 agggtcacgg ggtcagagga cgcaccttcc gaggggagga agctctctga ggccgcatca
     6241 tcttcagaac ttttgttcaa gaaacgtttg cgagcttgct ggaggagttc gtccttacgc
     6301 tgcaccagca tgcgctgtct ctcatcagca gacttggaga agcggctccc acgagcctcg
     6361 aagtcttcca cctcactggg ctccacttcc acctcgccga agtccagcgt ctcctccagg
     6421 cgaggactga ggtccagtgg cacacgctcg gtctgagcag aagtttctcc ctcttcctgg
     6481 tcactgcttg gcctttccac aggactgttc aatgcaggtc tgatgctatc tgaccgctgt
     6541 gtaggaaaag gtacttgaat ccgtccttct aaaatattgt ctgttgttat ttcaactgag
     6601 cgtgtcagct gaaggtcctg cagtaccaga tggtatggga cctggggaaa catctcttga
     6661 atctgatgag ccattgcatt gagctgggag ttgctggcct gcgtaatgcc aagaatgttg
     6721 gtggtgtgca tcacttcaac cgaaaaactc ggcagccagc tcgcaatccg agacccatcg
     6781 aaatggaaga agtgattgtg ttggtttaag cgaggtctcc cttcggctgc tgctacagga
     6841 accaaattct catccaagtt ctctccttga tgttcttccc tgacacgatt attgtcggca
     6901 atattaagag acattctgca tgttggacag gaggtgtctt gttctagcca ggaacgaaga
     6961 caggagttgt ggaaaagatg tccacagggc agtttccgcg cagcctgcat ggagccccaa
     7021 cagatggcac agtcgtcatt gttgacagcc agctcctctg gagttgcaac tgcaaacctg
     7081 gcctccatgt ttccaaccac acgtagacag ttcttgtgcc gacgaattcg acgttgcacc
     7141 tcatgaaaca ggtaacgcag ctgcataaag atgaccaggc tggccatgga taaccagatg
     7201 ttgccaaata acaacatgtg aatatggtgc atgaggtcca gggacaggag agtgagctcc
     7261 atgactaagt ctgtgtaata gacatacgtc ccctttcctt cccacgtccc ttcgtggttg
     7321 aggtcccaga ggtgaattac gtatcgtaaa atcacatgag cagtcctcac tgtcacaaga
     7381 agagactctg cagccatgaa agccaaggtg tgcattccgt gggtgtagcc ggtgatggag
     7441 cagacggccg ccagtccaca gcaggaaagc agcatggcaa ccaacaggga caggactc
//
