LOCUS       sequence-118887-        4775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4775)
  TITLE     Other Plasmids
REFERENCE   2  (bases 1 to 4775)
  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"
COMMENT     Sequence Label: V22 CS2+ RFPce
FEATURES             Location/Qualifiers
     source          1..4775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(94..96,97..99,100..768)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLF"
     primer_bind     complement(277..297)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     495..514
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     689..712
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    824..958
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(848..867)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     902..921
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1071..1091)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1071..1089)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1110..1126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1110..1126)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1123..1145)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1134..1150
                     /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(1158..1164,1165..1182,1183..1188))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1203..1224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1341..1358)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1512..2100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1592..1611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2271..3062,3063..3131))
                     /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     2894..2913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3132..3236)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3262..3717)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3399..3420)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3611..3630
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3791..4775
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttgatggtgc gctcctccaa gaacgtcatc
      121 aaggagttca tgcgcttcaa ggtgcgcatg gagggcaccg tgaacggcca cgagttcgag
      181 atcgagggcg agggcgaggg ccgcccctac gagggccaca acaccgtgaa gctgaaggtg
      241 accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc
      301 tccaaggtgt acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc
      361 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
      421 caggactcct ccctgcagga cggctgcttc atctacaagg tgaagttcat cggcgtgaac
      481 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag
      541 cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc acaaggccct gaagctgaag
      601 gacggcggcc actacctggt ggagttcaag tccatctaca tggccaagaa gcccgtgcag
      661 ctgcccggct actactacgt ggactccaag ctggacatca cctcccacaa cgaggactac
      721 accatcgtgg agcagtacga gcgcaccgag ggccgccacc acctgttcga ttcgaattca
      781 aggcctctcg agcctctaga actatagtga gtcgtattac gtagatccag acatgataag
      841 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
      901 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
      961 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     1021 attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc cctttagtga
     1081 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     1141 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     1201 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     1261 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1321 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     1381 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1441 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1501 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1561 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1621 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1681 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1741 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1801 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1861 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1921 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     1981 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2041 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2101 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2161 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2221 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     2281 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     2341 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2401 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2461 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     2521 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     2581 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     2641 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     2701 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     2761 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     2821 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     2881 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     2941 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3001 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3061 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     3121 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3181 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     3241 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3301 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3361 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3421 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3481 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3541 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3601 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3661 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     3721 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     3781 tacgccagtc gaccatagcc aattcaatat ggcgtatatg gactcatgcc aattcaatat
     3841 ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat
     3901 ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat
     3961 ggcggacttg gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag
     4021 gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc
     4081 ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc
     4141 aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat
     4201 atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat
     4261 atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata
     4321 tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat
     4381 tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca
     4441 cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct
     4501 attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa
     4561 cttggcaagt acattactat tggaaggacg ccagggtaca ttggcagtac tcccattgac
     4621 gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg
     4681 gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat
     4741 gggaggtcta tataagcaat gctcgtttag ggaac
//
