LOCUS       sequence-7756-ex        4915 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 4915)
  AUTHORS   Matsuda T, Cepko CL
  TITLE     Controlled expression of transgenes introduced by in vivo 
            electroporation.
  JOURNAL   Proc Natl Acad Sci U S A. 2007 Jan 16;104(3):1027-32. doi: 
            10.1073/pnas.0610155104
  PUBMED    17209010
REFERENCE   2  (bases 1 to 4915)
  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: "10.1073/pnas"; 
            journalName: "Proc Natl Acad Sci U S A"; date: "2007-01-16- 16"; 
            volume: "104"; issue: "3"; pages: "1027-32"
COMMENT     Sequence Label: pHes1-DsRed
FEATURES             Location/Qualifiers
     source          1..4915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1171..1180
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1177..1854
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(1357..1377)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1769..1792
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1939..1958)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2004..2059
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2058..2077)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2418..2434)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2418..2434)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2431..2453)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2442..2458
                     /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(2466..2472,2473..2490,2491..2496))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2511..2532
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2591..2786
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2637..2772
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2792..2926
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2829..2848)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2883..2902
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2994..3011)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3165..3753)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3245..3264)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3924..4715,4716..4784))
                     /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     4547..4566
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4785..4889)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact aggatctaca cgcacgcaca cacacacatc ctctgggccc
       61 cggccgcagc cgcggcggca ataaaacatc ctggcacgtg ctccgactcc aggcgcgccc
      121 tcgccggccg gctgatgtca aactgcagct cggctggtgg agctcttaaa gggctcgcgc
      181 gcggggcgcc gaggccgcct tggccgcccg cggggtgagg aggggagagc cccgactccc
      241 ttgcccgccg tctatccgta ttgccaactg ggagcctgcg gggagaaagg tagagagagt
      301 ggcggaggaa cgagcgcgcc cctcctgtcg cattgctatc aaagcccccc ttgttcattt
      361 tttttttcgt ttgaaattcc atttcctttc tgcccagtag gaaattattc tgattgtttt
      421 ctggagaagc gtgaggtcca aggttgcaat tccaagagag gaatgaatgg gctagaggga
      481 gacaagcatc caaaagacac cttgctccga aaaccctggc tgtgaaggca tttgggggga
      541 cggtaagggc atgtttagcg tgtggggttt agtatctccc caacctcatg agcagttttg
      601 actcccctaa acatacagag ttcgagcggg aattccggaa gacattagag taacacatcc
      661 tctttacctt gttccctcct tttttcaatc actaaatttt gtcttggcct atatctgttc
      721 aaaatatttt tcaaatgaac ttattataca aagtagttat attgcatgca gcaagaacaa
      781 taaaaaccaa aggcctggcc acaaaagaaa tagactagac taaaactaag caaagcccag
      841 aggaaagagt tagcaaaggg ttaaaatcct tttgattgac gttgtagcct ccggtgcccc
      901 gggctcaggc gcgcgccatt ggccgccaga ccttgtgcct agcggccaat gggggggcgc
      961 agtccacgag cggtgccgcg tgtctcttcc tcccattggc tgaaagttac tgtgggaaag
     1021 aaagtttggg aagtttcaca cgagccgttc gcgtgcagtc ccagatatat atagaggccg
     1081 ccagggcctg cggatcacac aggatctgga gctggtgctg ataacagcgg aatcccctgt
     1141 ctggtaccgc gggcccggga tccaccggtc gccaccatgg cctcctccga gaacgtcatc
     1201 accgagttca tgcgcttcaa ggtgcgcatg gagggcaccg tgaacggcca cgagttcgag
     1261 atcgagggcg agggcgaggg ccgcccctac gagggccaca acaccgtgaa gctgaaggtg
     1321 accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc
     1381 tccaaggtgt acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc
     1441 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggc gaccgtgacc
     1501 caggactcct ccctgcagga cggctgcttc atctacaagg tgaagttcat cggcgtgaac
     1561 ttcccctccg acggccccgt gatgcagaag aagaccatgg gctgggaggc ctccaccgag
     1621 cgcctgtacc cccgcgacgg cgtgctgaag ggcgagaccc acaaggccct gaagctgaag
     1681 gacggcggcc actacctggt ggagttcaag tccatctaca tggccaagaa gcccgtgcag
     1741 ctgcccggct actactacgt ggacgccaag ctggacatca cctcccacaa cgaggactac
     1801 accatcgtgg agcagtacga gcgcaccgag ggccgccacc acctgttcct gtagcggccg
     1861 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     1921 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     1981 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     2041 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     2101 gaatgagtat ttggtttaga gtttggcaac atatgccata tgctggctgc catgaacaaa
     2161 ggtggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     2221 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     2281 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     2341 gactactccc agtcatagct gtccctcttc tcttatgaag atccctcgac ctgcagccca
     2401 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2461 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2521 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2581 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2641 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     2701 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     2761 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     2821 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2881 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     2941 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3001 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3061 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3121 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3181 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3241 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3301 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     3361 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3421 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3481 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3541 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3601 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3661 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3721 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3781 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3841 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3901 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3961 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4021 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4081 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4141 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4201 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4261 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4321 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4381 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4441 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4501 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4561 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4621 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4681 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4741 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4801 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4861 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctg
//
