LOCUS       CMV_AcrIIC3_cpGR        6510 bp    DNA     circular SYN 06-AUG-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6510)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2025 from SnapGene 8.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        133..512
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        513..716
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        761..779
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             799..804
     CDS             805..981
                     /codon_start=1
                     /label=AcrIIC3
                     /translation="MAFKRAIIFTSFNGFEKVSRTEKRRLAKIINARVSIIDEYLRAKD
                     TNASLDGQYRAFLF"
     CDS             991..1203
                     /codon_start=1
                     /label=AncGR2 C176
                     /label=AncGR2\C176
                     /note="/vntifkey=21
                     after Rihtar et al. 2023 
                     ('>https://www.nature.com/articles/s41589-022-01136-x)"
                     /translation="NSSQNWQRFYQLTKLLDSMHEMVGGLLQFCFYTFVNKSLSVEFPE
                     MLAEIISNQLPKFNAGSVKPLLFHQK"
     CDS             1207..1251
                     /codon_start=1
                     /label=(GGS)5 Linker
                     /translation="GGSGGSGGSGGSGGS"
     CDS             1255..1779
                     /codon_start=1
                     /label=AncGR2N175
                     /note="/vntifkey=21
                     after Rihtar et al. 2023 
                     ('>https://www.nature.com/articles/s41589-022-01136-x)"
                     /translation="LISLLEVIEPEVLYSGYDSTLPDTSTRLMSTLNRLGGRQVVSAVK
                     WAKALPGFRNLHLDDQMTLLQYSWMSLMAFSLGWRSYKQSNGNMLCFAPDLVINEERMQ
                     LPYMYDQCQQMLKISSEFVRLQVSYDEYLCMKVLLLLSTVPKDGLKSQAVFDEIRMTYI
                     KELGKAIVKREG"
     CDS             1789..1965
                     /codon_start=1
                     /label=AcrIIC3
                     /translation="NDESPAMTEFLAKLKAFAESCTGISIDAWEIEESEYVRLPVERRD
                     FLAAANGKEIFKI"
     polyA_signal    2008..2232
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2278..2706
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2720..3049
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2900..3035
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3116..3910
                     /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    4084..4205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4254..4270)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4278..4294
                     /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(4302..4332)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4347..4368
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4656..5241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5412..6272)
                     /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(6273..6377)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
       61 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
      121 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      181 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      241 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      301 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      361 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      421 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      481 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      541 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      601 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      661 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct
      721 ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact cactataggg
      781 agacccaagc tggctagcgc caccatggct ttcaagagag ctattatatt tacgtccttt
      841 aatggttttg agaaggtctc tcgaacagag aagcgacgcc ttgccaagat aattaacgcg
      901 agggtctcaa ttattgatga gtatctcaga gccaaggata caaacgcgag cctggatggc
      961 caataccgag cctttctgtt tggtccgggg aacagcagcc agaactggca gaggttctac
     1021 cagctgacca agctgctgga cagcatgcac gagatggtgg gcggcctgct ccagttctgc
     1081 ttctacacct tcgtgaacaa gagcctgagc gtggagttcc ccgagatgct ggccgagatc
     1141 atcagcaacc agctgcctaa gtttaatgcc ggcagcgtga agcccctgct gttccaccag
     1201 aaggggggcg gaagcggcgg atccggcgga tctggcggat cgggcgggag tggactgatc
     1261 agcctgctgg aggtgatcga gcccgaggtg ctgtacagcg gctacgacag caccctgccc
     1321 gacaccagca ccaggctgat gagcaccctg aacaggctgg gcggcaggca ggtggtgagc
     1381 gccgtgaagt gggccaaggc cctgcccggc ttcaggaacc tgcacctgga cgaccagatg
     1441 accctgctgc agtacagctg gatgagcctg atggccttca gcctgggctg gaggagctac
     1501 aagcagagca acggcaacat gctgtgcttc gcccccgacc tggtgatcaa cgaggagagg
     1561 atgcagctgc cctacatgta cgaccagtgc cagcagatgc tgaagatcag cagcgagttc
     1621 gtgaggctgc aggtgagcta cgacgagtac ctgtgcatga aggtgctgct gctgctgagc
     1681 accgtgccca aggacggcct gaagagccag gccgtgttcg acgagatcag gatgacctac
     1741 atcaaggagc tgggcaaggc catcgtgaag agggagggcg gtccggggaa cgacgaaagc
     1801 cccgcgatga cagagttcct cgcaaaactg aaagcattcg cggagagctg cacgggaata
     1861 tcaatagatg cgtgggagat tgaagagagc gagtatgttc gactgcccgt tgaacgccgg
     1921 gatttcctgg ccgctgcgaa tggcaaggaa atcttcaaga tataactcga gcggccgcca
     1981 ctgtgctgga tatctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2041 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2101 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2161 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2221 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     2281 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2341 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2401 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2461 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2521 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2581 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2641 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2701 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2761 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     2821 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     2881 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     2941 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     3001 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     3061 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     3121 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3181 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3241 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3301 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3361 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3421 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3481 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3541 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3601 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     3661 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3721 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3781 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     3841 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     3901 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     3961 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4021 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4081 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4141 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4201 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     4261 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4321 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4381 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4441 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4501 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4561 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4621 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4681 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4741 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4801 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4861 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4921 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4981 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5041 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5101 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5161 tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg
     5221 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5281 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5341 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5401 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5461 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5521 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5581 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5641 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5701 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5761 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5821 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5881 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5941 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6001 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6061 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6121 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6181 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6241 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6301 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6361 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     6421 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     6481 gccagtatct gctccctgct tgtgtgttgg
//
