LOCUS       sequence-392246-        5218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AAV2 rep gene; accessory plasmid for the pCMV-Cap SSM 
            library..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5218)
  AUTHORS   Ogden PJ, Kelsic ED, Sinai S, Church GM
  TITLE     Comprehensive AAV capsid fitness landscape reveals a viral gene and 
            enables machine-guided design.
  JOURNAL   Science. 2019 Nov 29;366(6469):1139-1143. doi: 
            10.1126/science.aaw2900.
  PUBMED    31780559
REFERENCE   2  (bases 1 to 5218)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2019-11-29- 29"; volume: "366"; 
            issue: "6469"; pages: "1139-1143"
COMMENT     Sequence Label: pRep
FEATURES             Location/Qualifiers
     source          1..5218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     197..216
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(435..539)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(565..1020)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(702..723)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     914..933
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1147..1169
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1161..1178
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1162..1178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1188..1207
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1188..1206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1235..1251
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1236..1252
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3592..3612)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3592..3610)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3631..3647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3631..3647)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3644..3666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3655..3671
                     /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(3679..3685,3686..3703,3704..3709))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3724..3745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3862..3879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4033..4621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4113..4132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4792..5218,1..365,366..434))
                     /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"
ORIGIN
        1 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
       61 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
      121 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
      181 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
      241 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
      301 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
      361 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
      421 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
      481 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
      541 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
      601 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
      661 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
      721 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
      781 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
      841 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
      901 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
      961 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     1021 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     1081 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     1141 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
     1201 tatagggcga attgggtacc gggccccccc tcgatcgagg tcgacggtat cgggggagct
     1261 cgcagggtct ccattttgaa gcgggaggtt tgaacgcgca gccgccatgc cggggtttta
     1321 cgagattgtg attaaggtcc ccagcgacct tgacgagcat ctgcccggca tttctgacag
     1381 ctttgtgaac tgggtggccg agaaggaatg ggagttgccg ccagattctg acatggatct
     1441 gaatctgatt gagcaggcac ccctgaccgt ggccgagaag ctgcagcgcg actttctgac
     1501 ggaatggcgc cgtgtgagta aggccccgga ggctcttttc tttgtgcaat ttgagaaggg
     1561 agagagctac ttccacatgc acgtgctcgt ggaaaccacc ggggtgaaat ccatggtttt
     1621 gggacgtttc ctgagtcaga ttcgcgaaaa actgattcag agaatttacc gcgggatcga
     1681 gccgactttg ccaaactggt tcgcggtcac aaagaccaga aatggcgccg gaggcgggaa
     1741 caaggtggtg gatgagtgct acatccccaa ttacttgctc cccaaaaccc agcctgagct
     1801 ccagtgggcg tggactaata tggaacagta tttaagcgcc tgtttgaatc tcacggagcg
     1861 taaacggttg gtggcgcagc atctgacgca cgtgtcgcag acgcaggagc agaacaaaga
     1921 gaatcagaat cccaattctg atgcgccggt gatcagatca aaaacttcag ccaggtacat
     1981 ggagctggtc gggtggctcg tggacaaggg gattacctcg gagaagcagt ggatccagga
     2041 ggaccaggcc tcatacatct ccttcaatgc ggcctccaac tcgcggtccc aaatcaaggc
     2101 tgccttggac aatgcgggaa agattatgag cctgactaaa accgcccccg actacctggt
     2161 gggccagcag cccgtggagg acatttccag caatcggatt tataaaattt tggaactaaa
     2221 cgggtacgat ccccaatatg cggcttccgt ctttctggga tgggccacga aaaagttcgg
     2281 caagaggaac accatctggc tgtttgggcc tgcaactacc gggaagacca acatcgcgga
     2341 ggccatagcc cacactgtgc ccttctacgg gtgcgtaaac tggaccaatg agaactttcc
     2401 cttcaacgac tgtgtcgaca agatggtgat ctggtgggag gaggggaaga tgaccgccaa
     2461 ggtcgtggag tcggccaaag ccattctcgg aggaagcaag gtgcgcgtgg accagaaatg
     2521 caagtcctcg gcccagatag acccgactcc cgtgatcgtc acctccaaca ccaacatgtg
     2581 cgccgtgatt gacgggaact caacgacctt cgaacaccag cagccgttgc aagaccggat
     2641 gttcaaattt gaactcaccc gccgtctgga tcatgacttt gggaaggtca ccaagcagga
     2701 agtcaaagac tttttccggt gggcaaagga tcacgtggtt gaggtggagc atgaattcta
     2761 cgtcaaaaag ggtggagcca agaaaagacc cgcccccagt gacgcagata taagtgagcc
     2821 caaacgggtg cgcgagtcag ttgcgcagcc atcgacgtca gacgcggaag cttcgatcaa
     2881 ctacgcagac aggtaccaaa acaaatgttc tcgtcacgtg ggcatgaatc tgatgctgtt
     2941 tccctgcaga caatgcgaga gaatgaatca gaattcaaat atctgcttca ctcacggaca
     3001 gaaagactgt ttagagtgct ttcccgtgtc agaatctcaa cccgtttctg tcgtcaaaaa
     3061 ggcgtatcag aaactgtgct acattcatca tatcatggga aaggtgccag acgcttgcac
     3121 tgcctgcgat ctggtcaatg tggatttgga tgactgcatc tttgaacaat aaatgattta
     3181 aatcaggtat ggctgccgat ggttatcttc cagattggct cgaggacact ctctctgaag
     3241 gaataagaca gtggtggaag ctcaaacctg gcccaccatg ataataattg cctgttaatc
     3301 aataaaccgg ttgattcgtt tcagttgaac tttggtctct gcgaagggcg aattcgttta
     3361 aacctgcagg actagaggtc ctgtattaga ggtcacgtga gtgttttgcg acattttgcg
     3421 acaccatgtg gtcacgctgg gtatttaagc ccgagtgagc acgcagggtc tccattttga
     3481 agcgggaggt ttgaacgcgc agccgccaag ccgaattctg cagatatcca tcacactggc
     3541 ggccgctcga ctagagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg
     3601 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3661 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3721 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3781 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3841 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3901 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3961 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4021 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4081 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4141 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4201 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4261 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4321 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4381 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4441 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4501 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4561 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4621 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4681 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4741 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4801 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4861 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4921 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4981 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5041 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5101 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5161 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagc
//
