LOCUS       sequence-483016-        4616 bp    DNA     circular SYN 12-AUG-2025
DEFINITION  Template for in vitro transcription of mTurquoise2 mRNA using SP6 
            polymerase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4616)
  AUTHORS   n/a
  TITLE     Aiptasia Genetic Tools Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4616)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pCR175
FEATURES             Location/Qualifiers
     source          1..4616
                     /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             join(950..952,953..955,956..1669)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(995..1016)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1256..1275)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1603..1624
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1695..1714)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1696..1714)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1719..1853
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1743..1762)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1797..1816
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1970..1990)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1970..1988)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2009..2025)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2009..2025)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2022..2044)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2033..2049
                     /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(2057..2063,2064..2081,2082..2087))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2102..2123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2240..2257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2411..2999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2491..2510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3170..3961,3962..4030))
                     /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     3793..3812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4031..4135)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4161..4616)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4298..4319)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4510..4529
                     /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 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1021 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1081 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1141 cctgtcctgg ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     1201 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1261 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1321 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1381 caactacttt agcgacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     1441 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     1501 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1561 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1621 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaga attcaaggcc
     1681 tctcgagcct ctcgccctat agtgagtcgt attacgtaga tccagacatg ataagataca
     1741 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1801 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1861 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg
     1921 cggcgcgccg cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag
     1981 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2041 cgctcacaat tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct
     2101 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2161 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2221 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2281 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2341 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2401 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2461 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2521 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2581 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2641 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2701 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2761 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2821 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2881 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2941 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3001 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3061 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3121 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3181 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3241 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3301 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3361 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3421 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3481 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3541 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3601 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3661 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3721 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3781 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3841 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3901 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3961 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4021 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4081 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4141 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     4201 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4261 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     4321 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4381 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4441 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4501 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     4561 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgt
//
