LOCUS       sequence-464283-        7827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses SP1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7827)
  AUTHORS   Chou WC, Chen WT, Kuo CT, Chang YM, Lu YS, Li CW, Hung MC, Shen CY
  TITLE     Genetic insights into carbohydrate sulfotransferase 8 and its impact
            on the immunotherapy efficacy of cancer.
  JOURNAL   Cell Rep. 2024 Jan 23;43(1):113641. doi: 
            10.1016/j.celrep.2023.113641. Epub 2023 Dec 30.
  PUBMED    38165805
REFERENCE   2  (bases 1 to 7827)
  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: "Cell Rep.";
            date: "2024-01-23"; volume: "43(1):113641. doi"; pages: " 
            10.1016/j.celrep.2023.113641. Epub 2023 Dec 30"
COMMENT     Sequence Label: pcDNA3-Flag-SP1
FEATURES             Location/Qualifiers
     source          1..7827
                     /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             916..981
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(3380..3398)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3381..3398)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3418..3435)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3424..3648
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3694..4122
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3781..3800)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3991..4012
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4131..4151)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4136..4465
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4316..4451
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4378..4397
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4532..5326
                     /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(4586..4605)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5196..5215
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5500..5621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5537..5556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5591..5610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5670..5686)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5670..5686)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5683..5705)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5694..5710
                     /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(5718..5724,5725..5742,5743..5748))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5763..5784
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5901..5918)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6072..6660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6152..6171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6831..7622,7623..7691))
                     /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     7454..7473
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7692..7796)
                     /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 ccatggacta caaagaccat gacggtgatt ataaagatca tgacatcgac
      961 tacaaagacg atgacgacaa ggaattcgcc accatgagcg accaagatca ctccatggat
     1021 gaaatgacag ctgtggtgaa aattgaaaaa ggagttggtg gcaataatgg gggcaatggt
     1081 aatggtggtg gtgccttttc acaggctcga agtagcagca caggcagtag cagcagcact
     1141 ggaggaggag ggcaggagtc ccagccatcc cctttggctc tgctggcagc aacttgcagc
     1201 agaattgagt cacccaatga gaacagcaac aactcccagg gcccgagtca gtcaggggga
     1261 acaggtgagc ttgacctcac agccacacaa ctttcacagg gtgccaatgg ctggcagatc
     1321 atctcttcct cctctggggc tacccctacc tcaaaggaac agagtggcag cagtaccaat
     1381 ggcagcaatg gcagtgagtc ttccaagaat cgcacagtct ctggtgggca gtatgttgtg
     1441 gctgccgctc ccaacttaca gaaccagcaa gttctgacag gactacctgg agtgatgcct
     1501 aatattcagt atcaagtaat cccacagttc cagaccgttg atgggcaaca gctgcagttt
     1561 gctgccactg gggcccaagt gcagcaggat ggttctggtc aaatacagat cataccaggt
     1621 gcaaaccaac agattatcac aaatcgagga agtggaggca acatcattgc tgctatgcca
     1681 aacctactcc agcaggctgt ccccctccaa ggcctggcta ataatgtact ctcaggacag
     1741 actcagtatg tgaccaatgt accagtggcc ctgaatggga acatcacctt gctacctgtc
     1801 aacagcgttt ctgcagctac cttgactccc agctctcagg cagtcacgat cagcagctct
     1861 gggtcccagg agagtggctc acagcctgtc acctcaggga ctaccatcag ttctgccagc
     1921 ttggtatcat cacaagccag ttccagctcc tttttcacca atgccaatag ctactcaact
     1981 actactacca ccagcaacat gggaattatg aactttacta ccagtggatc atcagggacc
     2041 aactctcaag gccagacacc ccagagggtc agtgggctac aggggtctga tgctctgaac
     2101 atccagcaaa accagacatc tggaggctca ttgcaagcag gccagcaaaa agaaggagag
     2161 caaaaccagc agacacagca gcaacaaatt cttatccagc ctcagctagt tcaaggggga
     2221 caggccctcc aggccctcca agcagcacca ttgtcagggc agacctttac aactcaagcc
     2281 atctcccagg aaaccctcca gaacctccag cttcaggctg ttccaaactc tggtcccatc
     2341 atcatccgga caccaacagt ggggcccaat ggacaggtca gttggcagac tctacagctg
     2401 cagaacctcc aagttcagaa cccacaagcc caaacaatca ccttagcccc aatgcagggt
     2461 gtttccttgg ggcagaccag cagcagcaac accactctca cacccattgc ctcagctgct
     2521 tccattcctg ctggcacagt cactgtgaat gctgctcaac tctcctccat gccaggcctc
     2581 cagaccatta acctcagtgc attgggtact tcaggaatcc aggtgcaccc aattcaaggc
     2641 ctgccgttgg ctatagcaaa tgccccaggt gatcatggag ctcagcttgg tctccatggg
     2701 gctggtggtg atggaataca tgatgacaca gcaggtggag aggaaggaga aaacagccca
     2761 gatgcccaac cccaagccgg tcggaggacc cggcgggaag catgcacctg cccctactgt
     2821 aaagacagtg aaggaagggg ctcgggggat cctggcaaaa agaaacagca tatttgccac
     2881 atccaaggct gtgggaaagt gtatggcaag acctctcacc tgcgggcaca cttgcgctgg
     2941 catacaggcg agaggccatt tatgtgtacc tggtcatact gtgggaaacg cttcacacgt
     3001 tcggatgagc tacagaggca caaacgtaca cacacaggtg agaagaaatt tgcctgccct
     3061 gagtgtccta agcgcttcat gaggagtgac cacctgtcaa aacatatcaa gacccaccag
     3121 aataagaagg gaggcccagg tgtagctctg agtgtgggca ctttgcccct ggacagtggg
     3181 gcaggttcag aaggcagtgg cactgccact ccttcagccc ttattaccac caatatggta
     3241 gccatggagg ccatctgtcc agagggcatt gcccgtcttg ccaacagtgg catcaacgtc
     3301 atgcaggtgg cagatctgca gtccattaat atcagtggca atggcttctg agctcgagca
     3361 tgcatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
     3421 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     3481 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3541 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3601 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     3661 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3721 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3781 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3841 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3901 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3961 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4021 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4081 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     4141 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4201 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4261 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4321 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4381 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     4441 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     4501 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4561 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4621 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4681 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4741 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4801 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4861 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4921 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4981 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5041 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5101 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5161 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5221 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5281 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5341 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5401 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5461 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5521 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5581 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5641 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5701 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5761 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5821 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5881 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5941 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6001 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6061 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6121 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6181 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6241 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6301 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6361 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6421 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6481 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6541 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6601 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6661 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6721 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6781 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6841 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6901 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6961 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7021 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7081 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7141 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7201 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7261 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7321 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7381 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7441 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7501 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7561 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7621 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7681 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7741 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7801 tttccccgaa aagtgccacc tgacgtc
//
