LOCUS       pCR182                  4825 bp    DNA     circular SYN 04-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4825)
  AUTHORS   Christian Renicke
  TITLE     Direct Submission
  JOURNAL   Exported Jun 4, 2025 from SnapGene 8.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        78..1062
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1097..1115
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             1168..1878
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013).
                     
                     Codon-optimized for Exaiptasia diaphana."
                     /label=AmNeonGreen
                     /note="14 rare Aiptasia Pallida Transcriptome codons were 
                     removed."
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     promoter        complement(1905..1923)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1928..2062
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(2179..2197)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2218..2234)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2242..2258
                     /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(2266..2272,2273..2290,2291..2296))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2311..2332
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2620..3208)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3379..4170,4171..4239))
                     /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"
     promoter        complement(4240..4344)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4370..4825)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gtcgatcgac catagccaat tcaatatggc gtatatggac tcatgccaat
      121 tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat
      181 tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat
      241 tcaatatggc ggacttggca ctgtgccaac tggggagggg tctacttggc acggtgccaa
      301 gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa
      361 taatggcggc catattggct atatgccagg atcaatatat aggcaatatc caatatggcc
      421 ctatgccaat atggctattg gccaggttca atactatgta ttggccctat gccatatagt
      481 attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat
      541 accatatatg gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct
      601 ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct
      661 cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat
      721 aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac
      781 gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta
      841 atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc
      901 cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt
      961 caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt
     1021 gcctaatggg aggtctatat aagcaatgct cgtttaggga accgccattc tgcctgggga
     1081 cgtcggagca agcttgattt aggtgacact atagaataca agctacttgt tctttttgca
     1141 ggatcccatc gattcgaatt cgtcgacatg gtgagcaagg gcgaggagga taacatggcc
     1201 tctctcccag cgacacatga gttacacatc tttggctcca tcaacggtgt ggactttgac
     1261 atggtgggtc agggcaccgg caatccaaat gatggttatg aggagttaaa cctaaagtcc
     1321 accaagggtg acctccagtt ctccccctgg attctcgtcc ctcatatcgg gtatggcttc
     1381 catcagtacc taccctaccc tgacgggatg agtcctttcc aggccgctat ggtagatggc
     1441 tccggatacc aagtccatcg tacaatgcag tttgaagatg gtgcctccct tactgttaac
     1501 tacagataca cctacgaggg aagccacatc aaaggagagg cccaggtgaa ggggactggt
     1561 ttccctgctg acggtcctgt gatgaccaac agtctcaccg ctgcggactg gtgcaggagt
     1621 aagaagactt accccaacga caaaaccatc atcagtacct ttaagtggag ttacaccact
     1681 ggaaatggca agagataccg tagcactgcg aggaccacct acacctttgc taagccaatg
     1741 gcggctaact atctcaagaa ccagcctatg tacgtgttcc gtaagacgga actcaagcac
     1801 tccaagaccg agctcaactt caaggagtgg caaaaggcct ttaccgatgt gatgggcatg
     1861 gacgagctgt acaagtagaa ttcaaggcct ctcgagcctc tcgccctata gtgagtcgta
     1921 ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1981 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2041 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2101 gggggaggtg tgggaggttt tttaattcgc ggcgcgccgc ggcgccaatg cattgggccc
     2161 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     2221 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2281 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2341 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2401 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2461 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2521 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2581 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2641 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2701 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2761 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2821 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2881 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2941 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3001 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3061 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3121 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3181 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3241 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3301 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3361 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3421 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3481 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3541 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3601 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3661 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3721 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3781 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3841 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3901 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3961 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4021 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4081 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4141 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4201 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4261 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4321 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     4381 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4441 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4501 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4561 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4621 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4681 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4741 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4801 cttaatgcgc cgctacaggg cgcgt
//
