LOCUS       sequence-261688-        7507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EGFP reporter was removed and the selection marker was modified as 
            blasticidin in the construct of SHC003. This construct could be used
            for expression of a gene by lentiviral approach..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7507)
  AUTHORS   Wang S, Crisman L, Miller J, Datta I, Gulbranson DR, Tian Y, Yin Q, 
            Yu H, Shen J
  TITLE     Inducible Exoc7/Exo70 knockout reveals a critical role of the 
            exocyst in insulin-regulated GLUT4 exocytosis.
  JOURNAL   J Biol Chem. 2019 Dec 27;294(52):19988-19996. doi: 
            10.1074/jbc.RA119.010821. Epub 2019 Nov 18.
  PUBMED    31740584
REFERENCE   2  (bases 1 to 7507)
  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"; doi: 
            "10.1074/jbc.RA119.010821"; journalName: "J Biol Chem"; date: 
            "2019-12-27- 27"; volume: "294"; issue: "52"; pages: "19988-19996"
COMMENT     Sequence Label: SHC003BSD-DelGFP
FEATURES             Location/Qualifiers
     source          1..7507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      246..834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     735..754
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     988..1005
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1122..1143
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1158..1163,1164..1181,1182..1188)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1196..1212
                     /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     1201..1223
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1220..1236
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1220..1236
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1255..1275
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1257..1275
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1303..1529
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1530..1710
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1757..1882
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2375..2608
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2793..2837
                     /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        3135..3383
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3135..3155
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3306..3325
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        3446..3749
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3750..3953
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3904..3924
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    4170..4287
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4336..4846
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     4765..4783
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     LTR             5328..5561
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5633..5754
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5670..5689)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5724..5743
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5794..5929
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5949..5968)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5950..5968)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5978..5995)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5978..5994)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5987..6009)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6136..6591
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     6433..6454
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6617..6721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6722..6790,6791..7507,1..75)
                     /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(6940..6959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
       61 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
      121 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
      181 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
      241 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
      301 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
      361 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
      421 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      481 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      541 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      601 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      661 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      721 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      781 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      841 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
      901 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
      961 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     1021 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     1081 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     1141 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     1201 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     1261 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
     1321 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
     1381 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
     1441 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
     1501 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
     1561 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     1621 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     1681 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     1741 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     1801 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     1861 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     1921 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     1981 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     2041 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     2101 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     2161 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     2221 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     2281 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     2341 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     2401 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     2461 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     2521 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     2581 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     2641 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     2701 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     2761 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     2821 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     2881 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     2941 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     3001 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     3061 gatccattcg attagtgaac ggatctcgac ggtatcgatc acgagactag cctcgagcgg
     3121 ccgccccctt caccgagggc ctatttccca tgattccttc atatttgcat atacgataca
     3181 aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa
     3241 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
     3301 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
     3361 tcttgtggaa aggacgaaac accggtagtt attaatagta atcaattacg gggtcattag
     3421 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     3481 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     3541 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     3601 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     3661 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     3721 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     3781 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     3841 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     3901 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     3961 tgaaccgtca gatccgctag cgctaccgtc gacccccagc atcctgcaga acgggggccc
     4021 catgttcgcc ttccgccgcg tggaggagga tcacagcaac accgagctgg gcatcgtgga
     4081 gtaccagcac gccttcaaga ccccggatgc agatgccggt gaagaataaa attctcgacc
     4141 tcgagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac
     4201 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     4261 aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag agatccactt
     4321 tggccgcggc tcgagggggt tggggttgcg ccttttccaa ggcagccctg ggtttgcgca
     4381 gggacgcggc tgctctgggc gtggttccgg gaaacgcagc ggcgccgacc ctgggtctcg
     4441 cacattcttc acgtccgttc gcagcgtcac ccggatcttc gccgctaccc ttgtgggccc
     4501 cccggcgacg cttcctgctc cgcccctaag tcgggaaggt tccttgcggt tcgcggcgtg
     4561 ccggacgtga caaacggaag ccgcacgtct cactagtacc ctcgcagacg gacagcgcca
     4621 gggagcaatg gcagcgcgcc gaccgcgatg ggctgtggcc aatagcggct gctcagcagg
     4681 gcgcgccgag agcagcggcc gggaaggggc ggtgcgggag gcggggtgtg gggcggtagt
     4741 gtgggccctg ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc
     4801 ggcagtcggc tccctcgttg accgaatcac cgacctctct ccccaggggg atccctagcc
     4861 accatgcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
     4921 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
     4981 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
     5041 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
     5101 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
     5161 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
     5221 gaattgctgc cctctggtta tgtgtgggag ggctaaggta cctttaagac caatgactta
     5281 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     5341 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
     5401 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     5461 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     5521 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     5581 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     5641 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5701 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5761 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5821 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5881 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac
     5941 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
     6001 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     6061 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     6121 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     6181 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttattcc
     6241 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     6301 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     6361 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     6421 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     6481 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     6541 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     6601 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     6661 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     6721 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6781 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6841 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6901 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6961 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     7021 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     7081 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     7141 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     7201 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     7261 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     7321 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     7381 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     7441 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     7501 cacgacg
//
