LOCUS       SPLICa_ER-MT_WT         7265 bp    DNA     circular SYN 15-OCT-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7265)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 8, 2025 from SnapGene 8.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1438..1447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1999..2055
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     polyA_signal    2862..3086
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3132..3560
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3574..3903
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3754..3889
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3970..4764
                     /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"
     polyA_signal    4938..5059
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5108..5124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5132..5148
                     /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(5156..5162,5163..5180,5181..5186))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5201..5222
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5510..6098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6269..7060,7061..7129))
                     /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(7130..7234)
                     /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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcacc
      961 atggacggca aaagataccg gtgcacagtg agaacaacct acacctttgc caagcctatg
     1021 gccgctaatt acctgaagaa ccagcctatg tacgtgttca gaaagaccga gctgaagcac
     1081 agcaagacag agctgaattt caaggaatgg cagaaggccg tgaccgatgt gatggggatg
     1141 gatgagcttt ataaggtggt gggcggctct ggcggcacag gcatggtgtc caagggcgag
     1201 gaggacaaca tggcttctct gccagccacc cacgagctgc acattttcgg cagcatcaac
     1261 ggggttgatt ttgacatggt gggacagggc tctggcaacc ctaatggcgg ctacgaggaa
     1321 ctgaatctga aaagcaccat gggcgacctg caattttctc catggatcct ggtcccccac
     1381 atcggctacg gcttccacca gtacctgcct taccctgacg gcatgagccc cttccaggcc
     1441 gccatggtgg acggcagcgg ctaccaggtg cacagaacca tgcagttcga ggacggcgcc
     1501 agcctcaccg tcaactaccg ctacacatac gaggggagcc acatcatcgg cgaagcccag
     1561 gtggaaggca ccggatttcc tgctgatgga cctgtgatga ccaacagcat tacaggcacc
     1621 ggaggagatg gcggctccgg aggtggctct aagctgagag tgggcaccgg tggcgacggc
     1681 ggctctggcg gtggatctaa gctccgggtg ctggaaggaa ccggaggcga tggcggtagc
     1741 ggtggcggat caaagctgcg cgttttcctg gctctcccta tcatcatggt cgtggccttc
     1801 agcatgtgca tcatctgcct gctgatggcc ggcgacacct ggaccgagac actggcctac
     1861 gtgctgttct ggggcgtggc tagcatcggc accttcttca tcatcctgta caacggaaag
     1921 gatttcgtgg atgcccctag actggtgcag aaagaaaaga tcgacggaag cctgaaggaa
     1981 ctggcttgtg gcagcggcgc tacaaacttc agcctgctga aacaggccgg agacgtggaa
     2041 gaaaaccctg gccctagatc cgacattatg gtgggcagaa actccgccat cgccgccggc
     2101 gtgtgcggcg ccctgtttat cggctattgt atctacttcg atagaaagag acggagcgac
     2161 cccaacggca cagccagcag cgaggataac aacatggccg aattcatggt cgacagctct
     2221 agaaggaagt ggtacaagac cggccacgcc gtgcgggtgc tgcggcggct gagcagcggc
     2281 ctggactgga acagaagcac tatgacctac cccaacgaca agacaatcat ttctaccagc
     2341 aagtggtcct acaccacagg cgagagcgac cagctgaccg aggagcagat cgccgagatc
     2401 aaggaactgt tctctctgtt cgacaaggac ggggatggaa caatcaccac cagagagctg
     2461 ggcaccgtga tgagaagtct gggccaaaac cccaccgagg ccgaactgca ggacatgatc
     2521 aacgaggtgg acgccgaggg cgacggaaca atcgatttcc ccgagttcct ggccatgatg
     2581 gccagaaaga tgaaataccg ggacacagag gaggagctta gagaggctta tggcgtgttc
     2641 gacaaagacg gcaatagcta catctccgcc gcagagctgc ggcatatcgt gaccaacctg
     2701 ggagagaagc tgaccgacga agaagtggac gagatgctga gagaagccga tatcgatggc
     2761 gacggccaag tgaactacga ggaattcgtg cggatgatga cagccaaatg agcggccgct
     2821 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     2881 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2941 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3001 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3061 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3121 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3181 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3241 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3301 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3361 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3421 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3481 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3541 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3601 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3661 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3721 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3781 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3841 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3901 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3961 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4021 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4081 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4141 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4201 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4261 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4321 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4381 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4441 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4501 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4561 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4621 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4681 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4741 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4801 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4861 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4921 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4981 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5041 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5101 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5161 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5221 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5281 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5341 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5401 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5461 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5521 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5581 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5641 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5701 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5761 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5821 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5881 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5941 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6001 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6061 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6121 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6181 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6241 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6301 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6361 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6421 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6481 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6541 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6601 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6661 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6721 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6781 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6841 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6901 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6961 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7021 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7081 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7141 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7201 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7261 acgtc
//
