LOCUS       sequence-482340-        5654 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express amino acid 631-650 of AAA Mutant GRP78 preceded by a 
            secretory and sorting peptide of alpha-melanocyte-stimulating 
            hormone (SSP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5654)
  AUTHORS   Tseng CC, Zhang P, Lee AS
  TITLE     The COOH-Terminal Proline-Rich Region of GRP78 Is a Key Regulator of
            Its Cell Surface Expression and Viability of Tamoxifen-Resistant 
            Breast Cancer Cells.
  JOURNAL   Neoplasia. 2019 Aug;21(8):837-848. doi: 10.1016/j.neo.2019.05.008. 
            Epub 2019 Jul 12.
  PUBMED    31306849
REFERENCE   2  (bases 1 to 5654)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neo.2019.05.008"; journalName: "Neoplasia"; date: 
            "2019-08"; volume: "21"; issue: "8"; pages: "837-848"
COMMENT     Sequence Label: pcDNA3-P-SSP-AAA Mutant
FEATURES             Location/Qualifiers
     source          1..5654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1207..1225)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1208..1225)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1245..1262)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1251..1475
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1521..1949
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1608..1627)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1818..1839
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1958..1978)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1963..2292
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2143..2278
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2205..2224
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2359..3153
                     /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(2413..2432)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3023..3042
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3327..3448
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3364..3383)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3418..3437
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3497..3513)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3497..3513)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3510..3532)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3521..3537
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3545..3551,3552..3569,3570..3575))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3590..3611
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3728..3745)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3899..4487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3979..3998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4658..5449,5450..5518))
                     /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     5281..5300
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5519..5623)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccatgccgag atcgtgctgc agccgctcgg gggccctgtt gctggccttg
      961 ctgcttcagg cctccatgga agtgcgtggc tggtgcctgg agagcagcca gtgtcaggac
     1021 ctcaccacgg aaagcaacct gctggagtgc atccgggcct gcaagccccg cgagggcaag
     1081 cgcatcagca aactctatgg aagtgcaggc gctgccgcaa ctggtgaaga ggatacagca
     1141 gaataagaat tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg
     1201 ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc
     1261 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     1321 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     1381 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     1441 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     1501 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1561 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1621 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1681 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1741 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1801 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     1861 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     1921 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     1981 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2041 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2101 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2161 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2221 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2281 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     2341 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     2401 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     2461 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     2521 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     2581 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     2641 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     2701 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     2761 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     2821 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     2881 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     2941 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3001 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3061 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3121 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     3181 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     3241 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3301 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     3361 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3421 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     3481 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3541 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3601 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3661 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3721 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3781 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3841 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3901 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3961 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4021 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4081 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4141 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4201 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4261 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4321 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4381 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4441 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4501 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4561 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4621 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4681 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4741 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4801 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4861 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4921 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4981 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5041 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5101 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5161 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5221 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5281 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5341 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5401 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5461 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5521 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5581 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5641 tgccacctga cgtc
//
