LOCUS       sequence-346965-        4374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of anti-CRISPR AcrIIC1 mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4374)
  AUTHORS   Takasugi PR, Wang S, Truong KT, Drage EP, Kanishka SN, Higbee MA, 
            Bamidele N, Ojelabi O, Sontheimer EJ, Gagnon JA
  TITLE     Orthogonal CRISPR-Cas tools for genome editing, inhibition, and 
            CRISPR recording in zebrafish embryos.
  JOURNAL   Genetics. 2021 Nov 4. pii: 6420709. doi: 10.1093/genetics/iyab196.
  PUBMED    34735006
REFERENCE   2  (bases 1 to 4374)
  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: "Genetics. 
            2021 Nov 4. pii: 6420709. doi: 10.1093/genetics/iyab196."
COMMENT     Sequence Label: pCS2-AcrIIC1
FEATURES             Location/Qualifiers
     source          1..4374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        324..342
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     324..341
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             398..418
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    715..849
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(739..758)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     793..812
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(962..982)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(962..980)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1001..1017)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1001..1017)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1014..1036)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1025..1041
                     /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(1049..1055,1056..1073,1074..1079))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1094..1115
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1232..1249)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1403..1991)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1483..1502)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2162..2953,2954..3022))
                     /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     2785..2804
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3023..3127)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3153..3608)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3290..3311)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3502..3521
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(3679..4374,1..289)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata
       61 tctattaata gtaacttggc aagtacatta ctattggaag tacgccaggg tacattggca
      121 gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca
      181 ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg
      241 taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaacc gccattctgc
      301 ctggggacgt cggagcaagc ttgatttagg tgacactata gaatacaagc tacttgttct
      361 ttttgcagga tcccatcgat tcgaattcgc caccatgcca aaaaagaaga gaaaggtaga
      421 agaccccatg aataaaactt ataaaattgg aaaaaatgcc gggtatgatg gctgcggtct
      481 ttgtcttgcg gccatttctg aaaatgaagc tatcaaagtt aagtatttgc gcgacatttg
      541 tcctgattac gatggcgatg ataaagctga ggattggctg agatggggaa cggacagccg
      601 cgtcaaagca gccgctcttg aaatggagca gtacgcatat acgtcggttg gtatggcctc
      661 atgttgggag tttgttgaac tatgatctag aactatagtg agtcgtatta cgtagatcca
      721 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
      781 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
      841 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
      901 gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt
      961 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1021 gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag
     1081 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1141 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1201 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1261 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1321 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1381 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1441 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1501 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1561 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1621 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1681 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1741 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1801 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     1861 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1921 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1981 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2041 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2101 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2161 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2221 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2281 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2341 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2401 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     2461 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2521 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     2581 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     2641 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     2701 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     2761 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     2821 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     2881 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     2941 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3001 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3061 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3121 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     3181 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3241 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3301 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3361 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3421 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3481 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3541 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3601 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     3661 ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa
     3721 ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa
     3781 ttcaatatgg tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa
     3841 ttcaatatgg cggacctggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca
     3901 agtttgagga ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca
     3961 ataatggcgg ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc
     4021 cctatgccaa tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag
     4081 tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata
     4141 taccatatat ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc
     4201 tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc
     4261 tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa
     4321 taggaccacc caccattgac gtcaatggga tggctcattg cccattcata tccg
//
