LOCUS       Exported                9285 bp ds-DNA     circular SYN 19-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9285)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Aug 29, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9285
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    228..249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        264..294
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    302..318
                     /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)."
     primer_bind     326..342
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        363..381
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        409..635
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             636..816
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    863..988
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1481..1714
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1899..1943
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2210..2326
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2349..2652
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2653..2856
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      2933..2942
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2939..3604
                     /codon_start=1
                     /product="monomeric Keima-Red fluorescent protein, also 
                     known as mKeima"
                     /label=mKeima-Red
                     /translation="MVSVIAKQMTYKVYMSGTVNGHYFEVEGDGKGKPYEGEQTVKLTV
                     TKGGPLPFAWDILSPQLQYGSIPFTKYPEDIPDYFKQSFPEGYTWERSMNFEDGAVCTV
                     SNDSSIQGNCFIYNVKISGENFPPNGPVMQKKTQGWEPSTERLFARDGMLIGNDYMALK
                     LEGGGHYLCEFKSTYKAKKPVRMPGRHEIDRKLDVTSHNRDYTSVEQCEIAIARHSLLG
                     "
     misc_feature    3652..4240
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4123..4134)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4296..4795
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             4813..5838
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             6212..6445
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6517..6638
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6678..6813
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6834..6852)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6862..6878)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7020..7475
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7501..7605
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7606..8466
                     /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"
     rep_origin      8637..9225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
      361 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg
      421 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
      481 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
      541 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag
      601 atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct
      661 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
      721 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
      781 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
      841 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
      901 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
      961 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     1021 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     1081 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     1141 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     1201 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     1261 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     1321 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     1381 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     1441 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     1501 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     1561 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     1621 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     1681 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     1741 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     1801 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     1861 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     1921 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     1981 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     2041 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     2101 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     2161 gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact tttaaaagaa
     2221 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     2281 tacaaactaa agaattacaa aaacaaatta caaaattcaa aattttatcg ataagcttgg
     2341 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     2401 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     2461 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     2521 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     2581 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     2641 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     2701 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     2761 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     2821 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
     2881 tggagacgcc atccacgctg ttttgacctc catagaagac accgactcta gagccaccat
     2941 ggtgagtgtg atcgctaaac aaatgaccta caaggtttat atgtcaggca cggtcaatgg
     3001 acactacttt gaggtcgaag gcgatggaaa aggaaagcct tacgagggag agcagacagt
     3061 aaagctcact gtcaccaagg gtggacctct gccatttgct tgggatattt tatcaccaca
     3121 gcttcagtac ggaagcatac cattcaccaa gtaccctgaa gacatccctg attatttcaa
     3181 gcagtcattc cctgagggat atacatggga gaggagcatg aactttgaag atggtgcagt
     3241 gtgtactgtc agcaatgatt ccagcatcca aggcaactgt ttcatctaca atgtcaaaat
     3301 ctctggtgag aactttcctc ccaatggacc tgttatgcag aagaagacac agggctggga
     3361 acccagcact gagcgtctct ttgcacgaga tggaatgctg ataggaaacg attatatggc
     3421 tctgaagttg gaaggaggtg gtcactattt gtgtgaattt aaatctactt acaaggcaaa
     3481 gaagcctgtg aggatgccag ggcgccacga gattgaccgc aaactggatg taaccagtca
     3541 caacagggat tacacatctg ttgagcagtg tgaaatagcc attgcacgcc actctttgct
     3601 cggtggatcc gcgatcgcct gatggcaacg tggcgcgccc tcgaggtcga caatcaacct
     3661 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     3721 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3781 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3841 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3901 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3961 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     4021 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     4081 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     4141 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     4201 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattct gcagatatcc
     4261 ggactagtga tctctcgagg ttaacgaatt ctaccgggta ggggaggcgc ttttcccaag
     4321 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct
     4381 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg
     4441 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct
     4501 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga
     4561 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg
     4621 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg
     4681 cgggctcagg ggcggggcgg gcgcccgaag gccctccgga ggcccggcat tctgcacgct
     4741 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg
     4801 catcccgcca ccatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     4861 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     4921 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     4981 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa
     5041 gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     5101 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
     5161 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc
     5221 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     5281 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     5341 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     5401 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     5461 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     5521 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     5581 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     5641 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     5701 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc
     5761 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     5821 ccgagggcaa aggaatagag tagatgccga ccgaacaaga gctgatttcg agaacgcctc
     5881 agccagcaac tcgcgcgagc ctagcaaggc aaatgcgaga gaacggcctt acgcttggtg
     5941 gcacagttct cgtccacagt tcgctaagct cgctcggctg ggtcgcggga gggccggtcg
     6001 cagtgattca ggcccttctg gattgtgttg gtccccaggg cacgattgtc atgcccacgc
     6061 actcgggtga tctgactgat cccgcagatt ggagatcgcc gcccgtgcct gccgattggg
     6121 tgcagatccg tcgagggccc ggtaccttta agaccaatga cttacaaggc agctgtagat
     6181 cttagccact ttttaaaaga aaagggggga ctggaagggc tagctcactc ccaacgaaga
     6241 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     6301 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     6361 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     6421 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     6481 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     6541 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6601 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     6661 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6721 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     6781 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     6841 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     6901 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     6961 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     7021 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     7081 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     7141 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     7201 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     7261 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     7321 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     7381 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     7441 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     7501 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     7561 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     7621 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     7681 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     7741 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     7801 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     7861 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     7921 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     7981 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     8041 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     8101 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     8161 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     8221 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     8281 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     8341 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     8401 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     8461 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     8521 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     8581 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     8641 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     8701 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     8761 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     8821 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     8881 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     8941 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     9001 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     9061 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     9121 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     9181 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     9241 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatg
//
