LOCUS       sequence-431429-        6812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CROPseq, CMV-driven destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6812)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 6812)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: pRDA_789
FEATURES             Location/Qualifiers
     source          1..6812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      74..502
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(161..180)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     371..392
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(511..531)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        516..845
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      696..831
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     758..777
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        893..940
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             959..1333
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    1463..1584
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1500..1519)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1554..1573
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1633..1649)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1633..1649)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1646..1668)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1657..1673
                     /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(1681..1687,1688..1705,1706..1711))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1726..1747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1864..1881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2035..2623)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2115..2134)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2794..3585,3586..3654))
                     /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"
     primer_bind     3417..3436
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3655..3759)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3834..3853)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4025..4404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4406..4604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             4622..4802
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4849..4974
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5467..5700
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5885..5929
                     /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    6227..6344
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 atttatggac cccagtgaag acaccgtcct tctgaggcgg aaagaaccag ctggggctct
       61 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
      121 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
      181 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
      241 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
      301 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
      361 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
      421 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      481 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
      541 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      601 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
      661 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      721 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
      781 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      841 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
      901 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
      961 ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga
     1021 gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt
     1081 ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa
     1141 caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt
     1201 cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc
     1261 gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga
     1321 ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt
     1381 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     1441 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     1501 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     1561 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     1621 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1681 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     1741 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1801 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1861 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1921 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1981 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2041 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2101 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2161 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2221 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2281 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2341 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2401 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2461 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2521 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2581 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2641 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2701 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2761 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2821 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2881 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2941 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3001 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3061 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3121 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3181 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3241 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3301 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3361 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3421 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3481 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3541 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3601 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3661 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3721 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3781 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     3841 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     3901 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     3961 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     4021 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     4081 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     4141 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     4201 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     4261 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     4321 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     4381 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     4441 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     4501 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     4561 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac
     4621 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     4681 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     4741 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     4801 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     4861 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     4921 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     4981 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     5041 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     5101 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     5161 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     5221 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     5281 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     5341 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     5401 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     5461 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     5521 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     5581 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     5641 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     5701 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     5761 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     5821 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     5881 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     5941 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     6001 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     6061 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     6121 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
     6181 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     6241 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     6301 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     6361 acagcagaga tccagtttgg ttaattaagg taccgggaca cacgtcttca ccagtaattt
     6421 acgtgggaaa ggtgcttgtg tcccaacagg ctaggatata atgctgaaac cgtcccccaa
     6481 gcgttcaggg tggggtttgc tacaacttcc gagtccaatg tgtccgtgtt catgatatat
     6541 atgctcaagg gcgagaattg gacctagctt tcgtgttagt acgtagcatg gtgacacaag
     6601 cacagtagat cctgcccgcc gatcgtatat attaagttaa ttcttatgga atataataac
     6661 atgtggatgg ccagtggtcg gttgttacac gcctaccgcg atgctgaatg acccggacta
     6721 gagtggcgaa atttatggcg tgtgacccgt tatgctccat ttcggtcagt gggtcattgc
     6781 tagtagtcga ttgcattgcc attctccgag tg
//
