LOCUS       sequence-240841-        7376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for co-expression of CTLA4 and mgSrtA on cell membrane. For 
            proof-of-concept monitoring and labeling of CD80/86 positive cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7376)
  AUTHORS   Ge Y, Chen L, Liu S, Zhao J, Zhang H, Chen PR
  TITLE     Enzyme-Mediated Intercellular Proximity Labeling for Detecting 
            Cell-Cell Interactions.
  JOURNAL   J Am Chem Soc. 2019 Feb 6;141(5):1833-1837. doi: 
            10.1021/jacs.8b10286. Epub 2019 Jan 29.
  PUBMED    30676735
REFERENCE   2  (bases 1 to 7376)
  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: 
            "10.1021/jacs.8b10286"; journalName: "J Am Chem Soc"; date: 
            "2019-02-6- 6"; volume: "141"; issue: "5"; pages: "1833-1837"
COMMENT     Sequence Label: pcDNA3.1-CTLA4-mgSrtA
FEATURES             Location/Qualifiers
     source          1..7376
                     /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             917..1618
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(1050..1068)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1318..1337
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1512..1535
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1628..1684
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1745..1768
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2970..2987)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2976..3200
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3246..3674
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3333..3352)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3543..3564
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3683..3703)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3688..4017
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3868..4003
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3930..3949
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4084..4878
                     /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(4138..4157)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4748..4767
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5052..5173
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5089..5108)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5143..5162
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5222..5238)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5222..5238)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5235..5257)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5246..5262
                     /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(5270..5276,5277..5294,5295..5300))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5315..5336
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5453..5470)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5624..6209)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5704..5723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6380..7171,7172..7240))
                     /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     7003..7022
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7241..7345)
                     /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 gctggctagc
      901 gtttaaactt aagcttatgg tgagcaaggg cgaggaggtc atcaaagagt tcatgcgctt
      961 caaggtgcgc atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga
     1021 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct
     1081 gcccttcgcc tgggacatcc tgtcccccca gttcatgtac ggctccaagg cgtacgtgaa
     1141 gcaccccgcc gacatccccg attacaagaa gctgtccttc cccgagggct tcaagtggga
     1201 gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg acccaggact cctccctgca
     1261 ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc ccgacggccc
     1321 cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt acccccgcga
     1381 cggcgtgctg aagggcgaga tccaccaggc cctgaagctg aaggacggcg gccactacct
     1441 ggtggagttc aagacgatct acatggccaa gaagcccgtg caactgcccg gctactacta
     1501 cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     1561 cgagcgctcc gagggccgcc accacctgtt cctgtacggc atggacgagc tgtacaaggg
     1621 aagcggagct actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg
     1681 accggtaccg tgcacgctgc tcctgctgtt ggcggccgcc ctggctccga ctcagacccg
     1741 cgccgactac aaggacgacg acgacaaggg aagcggactg gagcaggcaa agccacagat
     1801 acccaaggac aagagcaagg tggcaggata catagagata ccagacgctg acataaaaga
     1861 gcccgtgtac cccggacccg ccacaagaga acaactgaac agaggagtgt ccttcgccaa
     1921 agaaaaccag agcctggacg accaaaacat cagcatcgcc ggccacacct tcatcggaag
     1981 acccaactac cagttcacca acctgaaggc cgccaaaaag ggaagcatgg tgtacttcaa
     2041 ggtgggcaac gaaaccagaa aatacaagat gacctccata agaaacgtga agcccaccgc
     2101 cgtgggcgtc ctggacgaac aaaagggaaa ggacaagcag ctgaccctga tcacctgcga
     2161 tgatctgaac agagaaaccg gcgtgtggga gaccagaaaa atcctggtgg ccacagaagt
     2221 gaagaccggt agcaccagcg gcagcggcaa gcccggcagc ggcgagggca gcaccaaggg
     2281 cagtactagt ggaagtggta aacctggaag tggagatggg agtacgaaag gaacgcgtat
     2341 acaggtgacc caaccttcag tggtgttggc tagcagccat ggtgtcgcca gctttccatg
     2401 tgaatattca ccatcacaca acactgatga ggtccgggtg actgtgctgc ggcagacaaa
     2461 tgaccaaatg actgaggtct gtgccacgac attcacagag aagaatacag tgggcttcct
     2521 agattacccc ttctgcagtg gtacctttaa tgaaagcaga gtgaacctca ccatccaagg
     2581 actgagagct gttgacacgg gactgtacct ctgcaaggtg gaactcatgt acccaccgcc
     2641 atactttgtg ggcatgggca acgggacgca gatttatgtc attgatccag aaccatgccc
     2701 ggattctgac ttcctccttt ggatccttgt cgcagttagc ttggggttgt ttttttacag
     2761 tttcctggtc actgctgttt ctttgagcaa gatgctaaag aaaagaagtc ctcttacaac
     2821 aggggtctat gtgaaaatgc ccccaacaga gccagaatgt gaaaagcaat ttcagcctta
     2881 ttttattccc atcaactgag aattctgcag atatccagca cagtggcggc cgctcgagtc
     2941 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3001 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3061 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3121 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3181 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3241 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3301 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3361 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3421 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3481 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3541 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3601 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3661 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3721 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3781 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3841 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3901 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3961 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4021 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4081 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4141 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4201 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4261 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4321 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4381 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4441 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4501 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4561 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4621 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4681 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4741 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4801 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4861 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4921 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4981 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5041 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5101 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5161 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5221 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5281 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5341 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5401 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5461 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5521 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5581 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5641 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5701 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5761 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5821 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5881 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5941 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6001 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6061 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6121 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     6181 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6241 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6301 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6361 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6421 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6481 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6541 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6601 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6661 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6721 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6781 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6841 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6901 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6961 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7021 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7081 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7141 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7201 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7261 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7321 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
