LOCUS       sequence-339961-        7150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CCL20 sender construct in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7150)
  AUTHORS   Cho KF, Gillespie SM, Kalogriopoulos NA, Quezada MA, Jacko M, Monje 
            M, Ting AY
  TITLE     A light-gated transcriptional recorder for detecting cell-cell 
            contacts.
  JOURNAL   Elife. 2022 Mar 21;11:e70881. doi: 10.7554/eLife.70881.
  PUBMED    35311648
REFERENCE   2  (bases 1 to 7150)
  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: "Elife."; 
            date: "2022-03-21"; volume: "11:e70881. doi"; pages: " 
            10.7554/eLife.70881"
COMMENT     Sequence Label: pCAG CAG CCL20-pre-mGRASP
FEATURES             Location/Qualifiers
     source          1..7150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2114..2203
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     intron          3072..3204
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3187..3206
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3213..3801
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3266..3286)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3684..3695)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3828..3845)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3834..3945
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4041..4061)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4041..4059)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4172..4191)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4237..4292
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4291..4310)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4653..4669)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4653..4669)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4666..4688)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4677..4693
                     /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(4701..4707,4708..4725,4726..4731))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4746..4767
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4826..5021
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4872..5007
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5027..5161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5064..5083)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5118..5137
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5229..5246)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5400..5988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5480..5499)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6159..6950,6951..7019))
                     /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     6782..6801
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7020..7124)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gccrgggggg gggggggggg gsggggggsg
      541 gggsggggsg rggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcggtaccat
     1741 gtgctgtacc aagagtttgc tcctggctgc tttgatgtca gtgctgctac tccacctctg
     1801 cggcgaatca gaagcagcaa gcaactttga ctgctgtctt ggatacacag accgtattct
     1861 tcatcctaaa tttattgtgg gcttcacacg gcagctggcc aatgaaggct gtgacatcaa
     1921 tgctatcatc tttcacacaa agaaaaagtt gtctgtgtgc gcaaatccaa aacagacttg
     1981 ggtgaaatat attgtgcgtc tcctcagtaa aaaagtcaag aacatgggta ccgggaacgg
     2041 aaatggcaat ggtaacggga acggaaatgg caatggtaac gggaatacta gtgttaacac
     2101 cggtgcggcc gtttacccat acgatgttcc tgactatgcg ggctatccct atgacgtccc
     2161 ggactatgca ggatcctatc catatgacgt tccagattac gctccggccg ccaccggtct
     2221 taaggtcgac tttcagaaag ctagctctat tgtttataag aaggaggggg aacaggtgga
     2281 gttcagtttc ccattggcct tcacagtcga gaagctcaca ggatcaggag agttgtggtg
     2341 gcaggccgaa agagcctcct ccagcaagag ttggatcact tttgatctga aaaacaaaga
     2401 agtgagcgtg aagagagtca cacaggaccc taagctgcag atgggcaaga agttgccttt
     2461 gcacttgaca ctcccccaag ccctgcccca gtatgctggc tccgggaacc tcaccctcgc
     2521 cttggaggct aagaccggaa agttgcatca ggaggtcaac ctcgtggtga tgagggcaac
     2581 acagctgcag aagaacctga cctgcgaagt gtggggtccc acctcaccta aactgatgct
     2641 gtctctgaag ctggagaata aggaggctaa agtcagtaaa agagagaagg ctgtgtgggt
     2701 ccttaacccg gaggcaggga tgtggcagtg tcttcttagc gactctggac aggtgctgct
     2761 ggagagcaac atcaaggtgc ttcctacttg gtcaacacca gtacagccga tggccctgat
     2821 cgtactcggg ggagtcgccg gcctgcttct gtttatcggc ctgggcatct tcttctgcgt
     2881 acgctacaag tatcgcaaca gggatgaggg atcttaccac gttgacgaat ctaggaacta
     2941 tatcagtaac agcgctcaga gtaatggcgc cgtggttaaa gagaaacagc cctccagcgc
     3001 caaatcagca aacaaaaaca agaaaaacaa ggataaagag tactacgtac ttaagtaact
     3061 cgaggccgca ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg
     3121 gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca
     3181 tccactttgc ctttctctcc acaggtgtcg acaatcaacc tctggattac aaaatttgtg
     3241 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3301 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3361 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     3421 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     3481 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3541 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3601 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     3661 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3721 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3781 ccctttgggc cgcctccccg cctggaattc gagctcggta cgatcagcct cgactgtgcc
     3841 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3901 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3961 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4021 caatagccca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtwrtmatkg
     4081 tyatmssykw wtycwstsct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     4141 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     4201 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     4261 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     4321 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     4381 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     4441 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     4501 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     4561 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     4621 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4681 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4741 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4801 ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc
     4861 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4921 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4981 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc
     5041 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5101 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     5161 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5221 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5281 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5341 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5401 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5461 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5521 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5581 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5641 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5701 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5761 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5821 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5881 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5941 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6001 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6061 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6121 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6181 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6241 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6301 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6361 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6421 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6481 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6541 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6601 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6661 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6721 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6781 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6841 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6901 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6961 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7021 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7081 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7141 gtgccacctg
//
