LOCUS       sequence-472962-        7538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CD44, a single-pass transmembrane protein containing 
            variable exons 3 to 10..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7538)
  AUTHORS   Tseng CC, Stanciauskas R, Zhang P, Woo D, Wu K, Kelly K, Gill PS, Yu
            M, Pinaud F, Lee AS
  TITLE     GRP78 regulates CD44v membrane homeostasis and cell spreading in 
            tamoxifen-resistant breast cancer.
  JOURNAL   Life Sci Alliance. 2019 Aug 15;2(4):e201900377. doi: 
            10.26508/lsa.201900377. Print 2019 Aug.
  PUBMED    31416894
REFERENCE   2  (bases 1 to 7538)
  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: "Life Sci 
            Alliance."; date: "2019-08-15"; volume: "2(4):e201900377. doi"; 
            pages: " 10.26508/lsa.201900377. Print 2019 Aug"
COMMENT     Sequence Label: CD44v-HA (pcDNA3)
FEATURES             Location/Qualifiers
     source          1..7538
                     /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"
     CDS             3001..3027
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3091..3109)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3092..3109)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3129..3146)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3135..3359
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3405..3833
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3492..3511)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3702..3723
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3842..3862)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3847..4176
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4027..4162
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4089..4108
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4243..5037
                     /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(4297..4316)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4907..4926
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5211..5332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5248..5267)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5302..5321
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5381..5397)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5381..5397)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5394..5416)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5405..5421
                     /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(5429..5435,5436..5453,5454..5459))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5474..5495
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5612..5629)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5783..6371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5863..5882)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6542..7333,7334..7402))
                     /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     7165..7184
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7403..7507)
                     /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 atggacaagt tttggtggca cgcagcctgg ggactctgcc tcgtgccgct gagcctggcg
      961 cagatcgatt tgaatataac ctgccgcttt gcaggtgtat tccacgtgga gaaaaatggt
     1021 cgctacagca tctctcggac ggaggccgct gacctctgca aggctttcaa tagcaccttg
     1081 cccacaatgg cccagatgga gaaagctctg agcatcggat ttgagacctg caggtatggg
     1141 ttcatagaag ggcacgtggt gattccccgg atccacccca actccatctg tgcagcaaac
     1201 aacacagggg tgtacatcct cacatccaac acctcccagt atgacacata ttgcttcaat
     1261 gcttcagctc cacctgaaga agattgtaca tcagtcacag acctgcccaa tgcctttgat
     1321 ggaccaatta ccataactat tgttaaccgt gatggcaccc gctatgtcca gaaaggagaa
     1381 tacagaacga atcctgaaga catctacccc agcaacccta ctgatgatga cgtgagcagc
     1441 ggctcctcca gtgaaaggag cagcacttca ggaggttaca tcttttacac cttttctact
     1501 gtacacccca tcccagacga agacagtccc tggatcaccg acagcacaga cagaatccct
     1561 gctaccagta cgtcttcaaa taccatctca gcaggctggg agccaaatga agaaaatgaa
     1621 gatgaaagag acagacacct cagtttttct ggatcaggca ttgatgatga tgaagatttt
     1681 atctccagca ccatttcaac cacaccacgg gcttttgacc acacaaaaca gaaccaggac
     1741 tggacccagt ggaacccaag ccattcaaat ccggaagtgc tacttcagac aaccacaagg
     1801 atgactgcag atgtagacag aaatggcacc actgcttatg aaggaaactg gaacccagaa
     1861 gcacaccctc ccctcattca ccatgagcat catgaggaag aagagacccc acattctaca
     1921 agcacaatcc aggcaactcc tagtagtaca acggaagaaa cagctaccca gaaggaacag
     1981 tggtttggca acagatggca tgagggatat cgccaaacac ccaaagaaga ctcccattcg
     2041 acaacaggga cagctgcagc ctcagctcat accagccatc caatgcaagg aaggacaaca
     2101 ccaagcccag aggacagttc ctggactgat ttcttcaacc caatctcaca ccccatggga
     2161 cgaggtcatc aagcaggaag aaggatggat atggactcca gtcatagtat aacgcttcag
     2221 cctactgcaa atccaaacac aggtttggtg gaagatttgg acaggacagg acctctttca
     2281 atgacaacgc agcagagtaa ttctcagagc ttctctacat cacatgaagg cttggaagaa
     2341 gataaagacc atccaacaac ttctactctg acatcaagca ataggaatga tgtcacaggt
     2401 ggaagaagag acccaaatca ttctgaaggc tcaactactt tactggaagg ttatacctct
     2461 cattacccac acacgaagga aagcaggacc ttcatcccag tgacctcagc taagactggg
     2521 tcctttggag ttactgcagt tactgttgga gattccaact ctaatgtcaa tcgttcctta
     2581 tcaggagacc aagacacatt ccaccccagt ggggggtccc ataccactca tggatctgaa
     2641 tcagatggac actcacatgg gagtcaagaa ggtggagcaa acacaacctc tggtcctata
     2701 aggacacccc aaattccaga atggctgatc atcttggcat ccctcttggc cttggctttg
     2761 attcttgcag tttgcattgc agtcaacagt cgaagaaggt gtgggcagaa gaaaaagcta
     2821 gtgatcaaca gtggcaatgg agctgtggag gacagaaagc caagtggact caacggagag
     2881 gccagcaagt ctcaggaaat ggtgcatttg gtgaacaagg agtcgtcaga aactccagac
     2941 cagtttatga cagctgatga gacaaggaac ctgcagaatg tggacatgaa gattggggtg
     3001 tacccatacg atgttccaga ttacgcttaa gaattctgca gatatccatc acactggcgg
     3061 ccgctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg ctagagctcg
     3121 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3181 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3241 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3301 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3361 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3421 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3481 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3541 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3601 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3661 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3721 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3781 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3841 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3901 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3961 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4021 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4081 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4141 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4201 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4261 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4321 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4381 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4441 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4501 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4561 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4621 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4681 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4741 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4801 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4861 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4921 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4981 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5041 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5101 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5161 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5221 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5281 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5341 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5401 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5461 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5521 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5581 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5641 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5701 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5761 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5821 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5881 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5941 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6001 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6061 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6121 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6181 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6241 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6301 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6361 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6421 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6481 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6541 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6601 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6661 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6721 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6781 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6841 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6901 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6961 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7021 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7081 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7141 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7201 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7261 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7321 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7381 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7441 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7501 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
