LOCUS       sequence-323888-        6902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shRNA targeting the coding sequences of GOLPH3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6902)
  AUTHORS   Sun X, Mahajan D, Chen B, Song Z, Lu L
  TITLE     A quantitative study of the Golgi retention of glycosyltransferases.
  JOURNAL   J Cell Sci. 2021 Oct 15;134(20). pii: 272560. doi: 
            10.1242/jcs.258564. Epub 2021 Oct 21.
  PUBMED    34533190
REFERENCE   2  (bases 1 to 6902)
  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: "J Cell 
            Sci."; date: "2021-10-15"; volume: "134(20). pii: 272560. doi"; 
            pages: " 10.1242/jcs.258564. Epub 2021 Oct 21"
COMMENT     Sequence Label: GOLPH3 shRNA
FEATURES             Location/Qualifiers
     source          1..6902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..536
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     455..473
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             558..1157
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(558..577)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1054..1074
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1285..1518
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1590..1711
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1627..1646)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1681..1700
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1751..1886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1906..1925)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1907..1925)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1935..1952)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1935..1951)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1944..1966)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2093..2548
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2180..2199)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2390..2411
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2574..2678
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2679..2747,2748..3539)
                     /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(2897..2916)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3710..4298
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4199..4218
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(4440..4445,4446..4463,4464..4470)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4478..4494
                     /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)."
     primer_bind     4483..4505
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4502..4518
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4502..4518
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4537..4557
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4539..4557
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4585..4811
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4812..4992
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5039..5164
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5657..5890
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6075..6119
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        6417..6665
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6417..6437
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6588..6607
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6762..6879
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 agatccactt tggccgcggc tcgagggggt tggggttgcg ccttttccaa ggcagccctg
       61 ggtttgcgca gggacgcggc tgctctgggc gtggttccgg gaaacgcagc ggcgccgacc
      121 ctgggtctcg cacattcttc acgtccgttc gcagcgtcac ccggatcttc gccgctaccc
      181 ttgtgggccc cccggcgacg cttcctgctc cgcccctaag tcgggaaggt tccttgcggt
      241 tcgcggcgtg ccggacgtga caaacggaag ccgcacgtct cactagtacc ctcgcagacg
      301 gacagcgcca gggagcaatg gcagcgcgcc gaccgcgatg ggctgtggcc aatagcggct
      361 gctcagcagg gcgcgccgag agcagcggcc gggaaggggc ggtgcgggag gcggggtgtg
      421 gggcggtagt gtgggccctg ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg
      481 agcgcacgtc ggcagtcggc tccctcgttg accgaatcac cgacctctct ccccaggggg
      541 atccaccgga gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
      601 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
      661 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
      721 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
      781 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
      841 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
      901 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
      961 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     1021 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     1081 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     1141 gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac
     1201 gaccccatgc atcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     1261 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     1321 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     1381 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     1441 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     1501 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     1561 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     1621 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     1681 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     1741 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     1801 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     1861 gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg
     1921 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     1981 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     2041 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     2101 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2161 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2221 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2281 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2341 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2401 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2461 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2521 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     2581 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2641 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2701 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2761 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2821 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2881 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     2941 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3001 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3061 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3121 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3181 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3241 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3301 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3361 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3421 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3481 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3541 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3601 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3661 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3721 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3781 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3841 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3901 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3961 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4021 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4081 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4141 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4201 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4261 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4321 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4381 attctgtgga taaccgtatt accgcctttg agtgagctaa ctcattaggc accccaggct
     4441 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     4501 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
     4561 caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca
     4621 tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat
     4681 tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat
     4741 ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc
     4801 gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     4861 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     4921 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     4981 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
     5041 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
     5101 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
     5161 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
     5221 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
     5281 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
     5341 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
     5401 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
     5461 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac
     5521 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     5581 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     5641 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     5701 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     5761 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     5821 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     5881 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     5941 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     6001 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     6061 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     6121 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     6181 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     6241 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     6301 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     6361 acggatctcg acggtatcga tcacgagact agcctcgagc ggccgccccc ttcaccgagg
     6421 gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt agagagataa
     6481 ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt
     6541 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc
     6601 ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa
     6661 acaccggaat tagcattgag aggaaggtct cgagaccttc ctctcaatgc taattttttt
     6721 gaattctcga cctcgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
     6781 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     6841 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
     6901 ag
//
