LOCUS       sequence-334283-        4365 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows for transcription of control mCerulean fluorophore half for 
            injection into zebrafish embryos for BiFC assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4365)
  AUTHORS   Don EK, Maschirow A, Radford RAW, Scherer NM, Vidal-Itriago A, Hogan
            A, Maurel C, Formella I, Stoddart JJ, Hall TE, Lee A, Shi B, Cole 
            NJ, Laird AS, Badrock AP, Chung RS, Morsch M
  TITLE     In vivo Validation of Bimolecular Fluorescence Complementation 
            (BiFC) to Investigate Aggregate Formation in Amyotrophic Lateral 
            Sclerosis (ALS).
  JOURNAL   Mol Neurobiol. 2021 May;58(5):2061-2074. doi: 
            10.1007/s12035-020-02238-0. Epub 2021 Jan 7.
  PUBMED    33415684
REFERENCE   2  (bases 1 to 4365)
  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: 
            "10.1007/s12035-020-02238-0"; journalName: "Mol Neurobiol"; date: 
            "2021-05"; volume: "58"; issue: "5"; pages: "2061-2074"
COMMENT     Sequence Label: pCS2plus-mCerulean 156-239-GGGS
FEATURES             Location/Qualifiers
     source          1..4365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      38..47
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(82..100)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(83..100)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(135..1119)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      1190..1645
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1277..1296)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1487..1508
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1671..1775
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1776..1844,1845..2636)
                     /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(1994..2013)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2807..3395
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3296..3315
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3549..3566
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3683..3704
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3719..3724,3725..3742,3743..3749)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3757..3773
                     /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     3762..3784
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3781..3797
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3781..3797
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3816..3836
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3818..3836
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    3949..4083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3986..4005)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4040..4059
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 atcttgaagt tggccttgat tccgttcttc tgcttgtcca tggtggcggc ggatcctgca
       61 aaaagaacaa gtagcttgta ttctatagtg tcacctaaat caagcttgct ccgacgtccc
      121 caggcagaat ggcggttccc taaacgagca ttgcttatat agacctccca ttaggcacgc
      181 ctaccgccca tttacgtcaa tggaacgccc atttgcgtca ttgcccctcc ccattgacgt
      241 caatggggat gtacttggca gccatcgcgg gccatttacc gccattgacg tcaatgggag
      301 tactgccaat gtaccctggc gtacttccaa tagtaatgta cttgccaagt tactattaat
      361 agatattgat gtactgccaa gtgggccatt taccgtcatt gacgtcaata gggggcgtga
      421 gaacggatat gaatgggcaa tgagccatcc cattgacgtc aatggtgggt ggtcctattg
      481 acgtcaatgg gcattgagcc aggcgggcca tttaccgtaa ttgacgtcaa tgggggaggc
      541 gccatatacg tcaataggac cgcccatatg acgtcaatag gaaagaccat atatagagac
      601 cattgacgtc aatgggggag tggctatggg cggtattagg aagccccata tatggtatat
      661 gggaccgccc attgggaggg gctatctacg tcaataggaa aacccatata tggaatacta
      721 tatggcatag ggccaataca tagtattgaa cctggccaat agccatattg gcatagggcc
      781 atattggata ttgcctatat attgatcctg gcatatagcc aatatggccg ccattattgg
      841 caccatgcca attcaatatg gcggacttgg cacagggcca agacccctcc tcaaacttgg
      901 caccgtgcca agtagacccc tccccagttg gcacagtgcc aggtccgcca tattgaattg
      961 gcatggtgcc aagtccgcca tattgaattg gctggggtcc agatccacca tattgaattg
     1021 gcacgagtcc atatacgcca tattgaattg gcacaggtcc agatccacca tattgaattg
     1081 gcatgagtcc atatacgcca tattgaattg gcggtcgact ggcgtaatag cgaagaggcc
     1141 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     1201 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     1261 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     1321 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     1381 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     1441 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     1501 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     1561 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     1621 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     1681 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1741 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1801 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1861 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1921 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1981 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2041 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2101 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2161 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     2221 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     2281 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     2341 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     2401 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     2461 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     2521 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     2581 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     2641 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2701 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2761 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2821 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2881 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2941 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3001 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3061 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3121 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3181 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3241 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3301 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3361 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3421 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3481 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3541 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     3601 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     3661 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     3721 tacactttat gctcccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     3781 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
     3841 aaaagctggg taccgggccc aatgcattgg cgccgcggcc gcgaattaaa aaacctccca
     3901 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     3961 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4021 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     4081 atctacgtaa tacgactcac tatagttcta gattagcttc ccccgccgct tccgcctccg
     4141 cttcctccgc ccttgtacag ctcgtccatt cccagggtga ttccagcggc ggtcacaaat
     4201 tccagcagca ccatgtggtc gcgcttctcg ttagggtcct tgctcagctt gctctgggtg
     4261 ctcaggtagt ggttgtctgg cagcagcaca ggtccatctc cgataggggt gttctgctgg
     4321 tagtggtcgg ccagctgcac gcttccgtcc tcgatgttgt ggcgg
//
