LOCUS       AcrVA1_cpGR2(GGS        6718 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 6718)
  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..6718
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        143..522
                     /note="CMV enhancer human cytomegalovirus immediate early 
                     enhancer"
     promoter        523..726
                     /note="CMV promoter human cytomegalovirus (CMV)
                     immediateearly promoter"
     promoter        771..789
                     /note="T7 promoter promoter for bacteriophage T7
                     RNApolymerase"
     CDS             809..1117
                     /codon_start=1
                     /label=AcrVA1
                     /translation="MYEAKERYAKKKMQENTKIDTLTDEQHDALAQLCAFRHKFHSNKD
                     SLFLSESAFSGEFSFEMQSDENSKLREVGLPTIEWSFYDNSHIPDDSFREWFNFANYS"
     CDS             1127..1339
                     /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             1343..1387
                     /codon_start=1
                     /label=(GGS)5 Linker
                     /translation="GGSGGSGGSGGSGGS"
     CDS             1391..1915
                     /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             1925..2119
                     /codon_start=1
                     /label=AcrVA1
                     /translation="ETIQEQGLELDLDDDETYELVYDELYTEAMGEYEELNQDIEKYLR
                     RIDEEHGTQYCPTGFARLR"
     polyA_signal    2226..2450
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2496..2924
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication;
                     arrowindicates direction of (+) strand synthesis"
     promoter        2938..3267
                     /note="SV40 promoter SV40 enhancer and early promoter"
     rep_origin      3118..3253
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3334..4128
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=aph(3')-II (or nptII)
                     /note="NeoR/KanR confers resistance to neomycin,
                     kanamycin,and G418 (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4302..4423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4472..4488)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4496..4512
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator The lac repressor binds to the
                     lacoperator to inhibit transcription in E. coli. This 
                     inhibition can be relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4520..4550)
                     /note="lac promoter promoter for the E. coli lac operon"
     protein_bind    4565..4586
                     /label=E. coli catabolite activator protein binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site CAP binding activates
                     transcriptionin the presence of cAMP."
     rep_origin      complement(4874..5459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5630..6490)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="AmpR confers resistance to ampicillin,carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6491..6595)
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
ORIGIN
        1 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
       61 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      121 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      181 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      241 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      301 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      361 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      421 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      481 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      541 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      601 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      661 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      721 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
      781 cactataggg agacccaagc tggctagcat gtatgaagct aaagagcgct atgccaagaa
      841 gaagatgcag gagaatacta agattgacac cttgacagat gaacagcatg atgctcttgc
      901 acagctttgc gctttcagac ataagttcca tagcaacaag gacagtctgt tcctgtctga
      961 gtctgctttc agcggggaat tctccttcga aatgcagtca gacgaaaata gcaaactgag
     1021 ggaggtggga ttgccaacaa tagaatggag cttctatgac aactcccaca ttcccgacga
     1081 ctctttccga gagtggttta actttgctaa ttacagcggt ccggggaaca gcagccagaa
     1141 ctggcagagg ttctaccagc tgaccaagct gctggacagc atgcacgaga tggtgggcgg
     1201 cctgctccag ttctgcttct acaccttcgt gaacaagagc ctgagcgtgg agttccccga
     1261 gatgctggcc gagatcatca gcaaccagct gcctaagttt aatgccggca gcgtgaagcc
     1321 cctgctgttc caccagaagg ggggcggaag cggcggatcc ggcggatctg gcggatcggg
     1381 cgggagtgga ctgatcagcc tgctggaggt gatcgagccc gaggtgctgt acagcggcta
     1441 cgacagcacc ctgcccgaca ccagcaccag gctgatgagc accctgaaca ggctgggcgg
     1501 caggcaggtg gtgagcgccg tgaagtgggc caaggccctg cccggcttca ggaacctgca
     1561 cctggacgac cagatgaccc tgctgcagta cagctggatg agcctgatgg ccttcagcct
     1621 gggctggagg agctacaagc agagcaacgg caacatgctg tgcttcgccc ccgacctggt
     1681 gatcaacgag gagaggatgc agctgcccta catgtacgac cagtgccagc agatgctgaa
     1741 gatcagcagc gagttcgtga ggctgcaggt gagctacgac gagtacctgt gcatgaaggt
     1801 gctgctgctg ctgagcaccg tgcccaagga cggcctgaag agccaggccg tgttcgacga
     1861 gatcaggatg acctacatca aggagctggg caaggccatc gtgaagaggg agggcggtcc
     1921 gggggaaacc atccaagagc agggactgga gctcgatctg gacgacgatg aaacgtacga
     1981 actcgtatac gacgaattgt atactgaggc catgggagaa tatgaagagc taaaccaaga
     2041 cattgagaag tatctgaggc gtattgacga agagcacggc actcagtatt gtccgacagg
     2101 gtttgcgagg cttcggtgac tcgagcggcc gccactgtgc tggatatctg cagaattcca
     2161 ccacactgga ctagtggatc cgagctcggt accaagctta agtttaaacc gctgatcagc
     2221 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2281 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2341 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2401 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2461 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     2521 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2581 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2641 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2701 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2761 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2821 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     2881 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     2941 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3001 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3061 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3121 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3181 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     3241 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     3301 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3361 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3421 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3481 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3541 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3601 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3661 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3721 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3781 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3841 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     3901 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3961 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4021 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4081 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4141 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4201 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4261 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4321 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4381 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4441 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4501 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4561 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4621 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4681 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4741 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4801 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4861 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4921 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4981 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5041 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5101 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5161 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5221 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5281 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5341 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5401 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5461 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5521 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5581 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5641 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5701 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5761 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5821 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5881 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5941 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6001 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6061 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6121 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6181 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6241 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6301 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6361 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6421 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6481 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6541 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6601 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
     6661 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgct
//
