LOCUS       CMV_AcrIIA5_AsLO        6259 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 6259)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2025 from SnapGene 8.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        224..603
                     /note="CMV enhancer human cytomegalovirus immediate early 
                     enhancer"
     promoter        604..807
                     /note="CMV promoter human cytomegalovirus (CMV)
                     immediateearly promoter"
     promoter        852..870
                     /label=T7 promoter
                     /note="T7 promoter promoter for bacteriophage T7
                     RNApolymerase"
     CDS             890..1741
                     /codon_start=1
                     /label=AcrIIA5
                     /translation="MAYGKSRYNSYRKRNFSISDNQRREYAKKMKELEQAFENLLATTL
                     ERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGRNCRFLQGPETDRATVRKIR
                     DAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQYFIGVQLDGTEHVRDAAER
                     EGVMLIKKTAENIDEAAKELDGWYLSSMKDSAYKDFGKYEIRLSNHSADNRYHDLENGR
                     LIVNVKASKLNFVDIIENKLGKIIEKIDTLDLDKYRFINATKLERDIKCYYKGYKTKKD
                     VI"
     CDS             1010..1438
                     /codon_start=1
                     /label=AsLOV2 hs
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQ
                     YFIGVQLDGTEHVRDAAEREGVMLIKKTAENIDEAAKEL"
     polyA_signal    1848..2072
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2118..2546
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication;
                     arrowindicates direction of (+) strand synthesis"
     promoter        2560..2889
                     /note="SV40 promoter SV40 enhancer and early promoter"
     rep_origin      2740..2875
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2956..3750
                     /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    3924..4045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4094..4110)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4118..4134
                     /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(4142..4172)
                     /note="lac promoter promoter for the E. coli lac operon"
     protein_bind    4187..4208
                     /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(4496..5081)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5252..6112)
                     /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(6113..6217)
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
ORIGIN
        1 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
       61 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
      121 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
      181 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
      241 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      301 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      361 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      421 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      481 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      541 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      601 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      661 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      721 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      781 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg
      841 cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagca tggcatacgg
      901 taaaagccga tataactctt accggaaacg aaattttagt atcagtgaca accagagaag
      961 agaatacgcc aaaaaaatga aggaattgga gcaagcgttt gagaatttgc ttgctacaac
     1021 tttggagcgg attgagaaga atttcgtcat aactgatccc aggttgcctg ataatcctat
     1081 aatattcgcc tccgatagtt ttcttcaact tacggagtac tccagagagg agatactggg
     1141 aaggaattgc agattcctcc aaggtccaga aactgataga gctactgtga ggaagatacg
     1201 ggatgccatt gacaatcaaa ctgaagtgac agtgcaactt atcaattaca ccaagtcagg
     1261 gaaaaaattc tggaacctgt tccatttgca acccatgagg gaccaaaagg gcgatgtgca
     1321 atatttcata ggcgttcaac ttgatggtac tgaacacgtc agagatgcgg cggagcgaga
     1381 aggagtgatg ctgattaaga aaacagcaga gaatatcgac gaagcggcga aagagctgga
     1441 cgggtggtac ctctcatcca tgaaggatag cgcctacaaa gacttcggaa agtacgagat
     1501 aagactgtca aaccacagtg cagacaatag gtatcacgac cttgaaaatg gtcgccttat
     1561 cgttaatgta aaggcttcaa aacttaattt cgttgatatt atagagaaca agctgggcaa
     1621 aattattgag aagatcgata cgcttgattt ggataagtac agattcatta acgctactaa
     1681 actggaaagg gatataaaat gttactataa ggggtataaa acaaaaaagg atgttatctg
     1741 actcgagcgg ccgccactgt gctggatatc tgcagaattc caccacactg gactagtgga
     1801 tccgagctcg gtaccaagct taagtttaaa ccgctgatca gcctcgactg tgccttctag
     1861 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1921 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1981 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2041 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     2101 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2161 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2221 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2281 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2341 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2401 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2461 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2521 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     2581 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2641 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2701 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     2761 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     2821 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     2881 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     2941 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3001 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3061 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3121 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3181 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3241 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3301 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3361 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3421 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3481 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3541 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3601 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3661 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3721 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3781 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     3841 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     3901 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     3961 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4021 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4081 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4141 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4201 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4261 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4321 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4381 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4441 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4501 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4561 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4621 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4681 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4741 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4801 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4861 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4921 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4981 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     5041 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5101 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5161 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5221 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5281 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5341 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5401 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5461 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5521 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5581 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5641 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5701 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5761 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5821 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5881 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5941 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6001 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6061 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6121 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6181 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6241 ctgacgtcga cggatcggg
//
