LOCUS       sequence-60627-e        7340 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 7340)
  AUTHORS   Martell JD, Deerinck TJ, Sancak Y, Poulos TL, Mootha VK, Sosinsky 
            GE, Ellisman MH, Ting AY
  TITLE     Engineered ascorbate peroxidase as a genetically encoded reporter 
            for electron microscopy.
  JOURNAL   Nat Biotechnol. 2012 Oct 21. doi: 10.1038/nbt.2375.
  PUBMED    23086203
REFERENCE   2  (bases 1 to 7340)
  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"; journalName: "Nat 
            Biotechnol. 2012 Oct 21. doi: 10.1038/nbt.2375."
COMMENT     Sequence Label: pcDNA3 Connexin43-APEX
FEATURES             Location/Qualifiers
     source          1..7340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      917..926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2864..2893
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2934..2951)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2940..3164
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3210..3638
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3297..3316)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3507..3528
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3647..3667)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3652..3981
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3832..3967
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3894..3913
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4048..4842
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4102..4121)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4712..4731
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5016..5137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5053..5072)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5107..5126
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5186..5202)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5186..5202)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5199..5221)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5210..5226
                     /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(5234..5240,5241..5258,5259..5264))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5279..5300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5417..5434)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5588..6173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5668..5687)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6344..7135,7136..7204))
                     /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     6967..6986
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7205..7309)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatgggtga ctggagtgcc ttggggaaat tactggacaa
      961 ggtccaagcc tactccaccg ctggagggaa ggtgtggctg tcagtgctct tcatattcag
     1021 aatcctgctc ctggggacag ctgttgagtc agcttggggt gatgaacagt ctgcctttcg
     1081 ctgtaacact caacaacctg gctgcgaaaa cgtctgctat gacaagtcct tccccatctc
     1141 tcacgtgcgc ttctgggtcc ttcagatcat attcgtgtct gtgcccacac tcctgtactt
     1201 ggcccatgtg ttctatgtga tgaggaagga agagaagcta aacaagaaag aagaggagct
     1261 caaagtggcc cagactgacg gggtcaacgt ggagatgcac ctgaagcaga ttgaaatcaa
     1321 gaagttcaag tacgggattg aagagcacgg caaggtgaaa atgaggggcg gcttgctgag
     1381 aacctacatc atcagcatcc tcttcaagtc tgtcttcgag gtggccttcc tgctcatcca
     1441 gtggtacatc tatgggttca gcttgagcgc ggtctacacc tgcaagagag atccctgccc
     1501 ccaccaggta gactgcttcc tctcacgtcc cacggagaaa accatcttca tcatcttcat
     1561 gctggtggtg tccttggtgt ctctcgcttt gaacatcatt gagctcttct acgtcttctt
     1621 caagggcgtt aaggatcgcg tgaagggaag aagcgatcct taccacgcca ccactggccc
     1681 actgagccca tcaaaagact gcggatctcc aaaatacgcc tacttcaatg gctgctcctc
     1741 accaacggct ccactctcgc ctatgtctcc tcctgggtac aagctggtta ctggtgacag
     1801 aaacaattcc tcgtgccgca attacaacaa gcaagctagc gagcaaaact gggcgaacta
     1861 cagcgcagag caaaatcgca tggggcaggc cggaagcacc atctccaact cgcacgccca
     1921 gccgttcgat ttccccgacg acaaccagaa tgccaaaaaa gttgctgctg gacatgaact
     1981 ccagccatta gccatcgtgg accaacgacc ttccagcaga gccagcagcc gcgccagcag
     2041 caggcctcgg cctgatgacc tggagattgg atccaagggc tcgggctcga cctcgggctc
     2101 gggcagggga aaatcttacc caactgttag tcccgattac caggacgcca ttgaaaaggc
     2161 taagaggaag ctcagaggtt ttatcgctga gaagaaatgc gctcctctaa ttctccgttt
     2221 ggcattccac tctgctggta cttttgattc caagacaaag actggtggtc ctttcggaac
     2281 aattaagcac caagctgagc ttgctcatgg tgctaacaac ggtcttgata tcgcggttag
     2341 gctgttggag cctattaagg agcaattccc tattgtgagc tatgctgatt tctaccagtt
     2401 ggctggtgtt gttgctgttg agattaccgg tggacctaag gttcctttcc accctggtag
     2461 ggaggacaag cctgagccac cacctgaggg tcgcttgcct gatgccacta agggttctga
     2521 ccatttgagg gatgtgtttg gaaaggctat ggggcttagt gatcaggaca ttgttgctct
     2581 atctggtggt cacaccattg gagctgcaca caaggagcgt tctggatttg agggaccgtg
     2641 gacttctaat cctctcattt ttgacaactc atatttcact gagttgttga ctggtgagaa
     2701 ggatggcctt cttcagttgc caagtgataa ggcacttttg actgactctg tattccgccc
     2761 tcttgttgag aaatatgctg cggatgaaga tgttttcttt gctgattatg ctgaagcaca
     2821 tcttaagctc tctgagcttg gatttgctga agccggcggc cgcgaacaaa aacttatttc
     2881 tgaagaagat ctgtaactcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     2941 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3001 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3061 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3121 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3181 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3241 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3301 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3361 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3421 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3481 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3541 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3601 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3661 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3721 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3781 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3841 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3901 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3961 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4021 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4081 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4141 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4201 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4261 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4321 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4381 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4441 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4501 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4561 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4621 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4681 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4741 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4801 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4861 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4921 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4981 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5041 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5101 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5161 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5221 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5281 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5341 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5401 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5461 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5521 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5581 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5641 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5701 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5761 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5821 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5881 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5941 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6001 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6061 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6121 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6181 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6241 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6301 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6361 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6421 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6481 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6541 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6601 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6661 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6721 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6781 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6841 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6901 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6961 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7021 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7081 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7141 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7201 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7261 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7321 gaaaagtgcc acctgacgtc
//
