LOCUS       sequence-482342-        4595 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Template for in vitro transcription of mScarlet-I mRNA using SP6 
            polymerase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4595)
  AUTHORS   n/a
  TITLE     Aiptasia Genetic Tools Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4595)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pCR178
FEATURES             Location/Qualifiers
     source          1..4595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        879..897
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     879..896
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             950..1645
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1083..1101)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1351..1370
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1674..1693)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1675..1693)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1698..1832
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1722..1741)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1776..1795
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1949..1969)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1949..1967)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1988..2004)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1988..2004)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2001..2023)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2012..2028
                     /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(2036..2042,2043..2060,2061..2066))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2081..2102
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2219..2236)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2390..2978)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2470..2489)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3149..3940,3941..4009))
                     /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     3772..3791
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4010..4114)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4140..4595)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4277..4298)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4489..4508
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gtcgatcgac cgccaattca atatggcgta tatggactca tgccaattca
      121 atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca
      181 atatggtgga tctggacccc agccaattca atatggctat tggccaggtt caatactatg
      241 tattggccct atgccatata gtattccata tatgggtttt cctattgacg tagatagccc
      301 ctcccaatgg gcggtcccat ataccatata tggggcttcc taataccgcc catagccact
      361 cccccattga cgtcaatggt ctctatatat ggtctttcct attgacgtca tatgggcggt
      421 cctattgacg tatatggcgc ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg
      481 ctcaatgccc attgacgtca ataggaccac ccaccattga cgtcaatggg atggctcatt
      541 gcccattcat atccgttctc acgcccccta ttgacgtcaa tgacggtaaa tggcccactt
      601 ggcagtacat caatatctat taatagtaac ttggcaagta cattactatt ggaagtacgc
      661 cagggtacat tggcagtact cccattgacg tcaatggcgg taaatggccc gcgatggctg
      721 ccaagtacat ccccattgac gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt
      781 gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg
      841 gaaccgccat tctgcctggg gacgtcggag caagcttgat ttaggtgaca ctatagaata
      901 caagctactt gttctttttg caggatccca tcgattcgaa ttcgtcgaca tggtgagcaa
      961 gggcgaggca gtgatcaagg agttcatgcg gttcaaggtg cacatggagg gctccatgaa
     1021 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     1081 cgccaagctg aaggtgacca agggtggccc cctgcccttc tcctgggaca tcctgtcccc
     1141 tcagttcatg tacggctcca gggccttcat caagcacccc gccgacatcc ccgactacta
     1201 taagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     1261 cgccgtgacc gtgacccagg acacctccct ggaggacggc accctgatct acaaggtgaa
     1321 gctccgcggc accaacttcc ctcctgacgg ccccgtaatg cagaagaaga caatgggctg
     1381 ggaagcgtcc accgagcggt tgtaccccga ggacggcgtg ctgaagggcg acattaagat
     1441 ggccctgcgc ctgaaggacg gcggccgcta cctggcggac ttcaagacca cctacaaggc
     1501 caagaagccc gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg acatcacctc
     1561 ccacaacgag gactacaccg tggtggaaca gtacgaacgc tccgagggcc gccactccac
     1621 cggcggcatg gacgagctgt acaagtagaa ttcaaggcct ctcgagcctc tcgccctata
     1681 gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca
     1741 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     1801 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     1861 ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggcgcgccgc ggcgccaatg
     1921 cattgggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     1981 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     2041 tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2101 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2161 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2221 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2281 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2341 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2401 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2461 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2521 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2581 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2641 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2701 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2761 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2821 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2881 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2941 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3001 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3061 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3121 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3181 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3241 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3301 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3361 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3421 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3481 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3541 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3601 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3661 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3721 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3781 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3841 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3901 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3961 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4021 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4081 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     4141 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     4201 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     4261 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     4321 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     4381 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     4441 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     4501 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     4561 acccgccgcg cttaatgcgc cgctacaggg cgcgt
//
