LOCUS       pRK5R-MCS+              4674 bp ds-DNA     circular     02-MAY-2023
DEFINITION  .
FEATURES             Location/Qualifiers
     rep_origin      complement(1..589)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
                     /direction="RIGHT"
     primer_bind     complement(81..100)
                     /label="pBR322ori-F"
                     /note="pBR322 origin, forward primer"
     CDS             complement(760..1620)
                     /label="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, andrelated antibiotics"
                     /gene="bla"
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     primer_bind     1383..1402
                     /label="Amp-R"
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1621..1725)
                     /label="AmpR promoter"
                     /gene="bla"
     primer_bind     1793..1811
                     /label="pBRforEco"
                     /note="pBR322 vectors, upsteam of EcoRI site, forward primer"
     primer_bind     complement(1849..1871)
                     /label="pGEX 3'"
                     /note="pGEX vectors, reverse primer"
     primer_bind     1971..1990
                     /label="pRS-marker"
                     /note="pRS vectors, use to sequence yeast selectable marker"
     rep_origin      complement(2007..2462)
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
                     /direction="RIGHT"
     primer_bind     complement(2144..2165)
                     /label="F1ori-F"
                     /note="F1 origin, forward primer"
     primer_bind     2356..2375
                     /label="F1ori-R"
                     /note="F1 origin, reverse primer"
     primer_bind     2660..2682
                     /label="M13/pUC Forward"
                     /note="In lacZ gene"
     primer          complement(2672..2691)
                     /label="oJY92"
                     /note="sequence: ACTGGCCGTCGTTTTACAAC"
     primer          complement(2672..2691)
                     /label="oJY34"
                     /note="sequence: cgggcgagctcgaattaattACTGGCCGTCGTTTTACAAC"
     primer_bind     2674..2691
                     /label="M13 Forward"
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward"
     primer_bind     2675..2691
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar variants"
     primer          2692..2712
                     /label="oJY36"
                     /note="sequence: AATTAATTCGAGCTCGCCCGA"
     misc_feature    2692..3609
                     /label="frag 2"
     enhancer        2711..3090
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3091..3294
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early promoter"
     primer_bind     3245..3265
                     /label="CMV-F"
                     /note="Human CMV immediate early promoter, forward primer"
     primer          3286..3304
                     /label="RC426"
                     /note="sequence: agcagagctcgtttagtga"
     primer_bind     3291..3315
                     /label="LNCX"
                     /note="Human CMV promoter, forward primer"
     primer_bind     3525..3542
                     /label="SP6"
                     /note="SP6 promoter, forward primer"
     promoter        3525..3543
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3556..3575
                     /label="pMT2-F"
                     /note="Synthetic intron, forward primer"
     primer          complement(3590..3609)
                     /label="oJY37"
                     /note="sequence: TAGAACCGAGGTGCAGTTGG"
     misc_feature    3610..4243
                     /label="frag 3"
     primer          3662..3677
                     /label="oJY39"
                     /note="sequence: ccaactgcacctcggttctaggatccttagcggccgccgagaattcgaattaattaaaccggtagcgctagcCCAGCTTGGCCGCCAT"
     primer          3662..3677
                     /label="oJY38"
                     /note="sequence: ccaactgcacctcggttctagctagcgctaccggtttaattaattcgaattctcggcggccgctaaggatccCCAGCTTGGCCGCCAT"
     polyA_signal    3683..3817
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3720..3739)
                     /label="SV40pA-R"
                     /note="SV40 polyA, reverse primer"
     primer_bind     3774..3793
                     /label="EBV-rev"
                     /note="SV40 polyA terminator, reverse primer"
     primer          3836..3856
                     /label="oJY91"
                     /note="sequence: gcaccatggcctgaaataacc"
     promoter        3886..4243
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3909..3929)
                     /label="pBABE 3'"
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4094..4229
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     4156..4175
                     /label="SV40pro-F"
                     /note="SV40 promoter/origin, forward primer"
     primer          4182..4201
                     /label="oJY41"
                     /note="sequence: cggcctctgagctattccag"
     primer          complement(4223..4243)
                     /label="oJY40"
                     /note="sequence: TTTGCAAAAGCCTAGGCCTCC"
     misc_feature    4244..2691
                     /label="frag 1"
     primer          4263..4287
                     /label="oJY35"
                     /note="sequence: gaggcctaggcttttgcaaaaagctgttaacagcttggcTCGTAATCATGTCATAGCTGTTTCC"
     primer_bind     complement(4273..4289)
                     /label="M13 Reverse"
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4273..4289)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar variants"
     primer_bind     complement(4286..4308)
                     /label="M13/pUC Reverse"
                     /note="In lacZ gene"
     protein_bind    complement(4297..4313)
                     /label="lac operator"
                     /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)."
                     /bound_moiety="lac repressor encoded by lacI"
     promoter        complement(4321..4351)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4366..4387)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence of cAMP."
                     /bound_moiety="E. coli catabolite activator protein"
     primer_bind     complement(4504..4521)
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     primer          complement(4635..4654)
                     /label="RC457"
                     /note="sequence: tttacggttcctggcctttt"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1741 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     1801 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     1861 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     1921 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     1981 gcagattgta ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc
     2041 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2101 ttataaatca aaagaatagc ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2161 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2221 tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc
     2281 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2341 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2401 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2461 gtactatggt tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     2521 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     2581 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     2641 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag taattaattc
     2701 gagctcgccc gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     2761 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     2821 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     2881 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     2941 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     3001 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     3061 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     3121 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     3181 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     3241 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
     3301 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     3361 cgggaccgat ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc
     3421 aagagtgacg taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa
     3481 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat
     3541 agaataacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc
     3601 tcggttctag ctagcgctac cggtttaatt aattcgaatt ctcggcggcc gctaaggatc
     3661 cccagcttgg ccgccatggc ccaacttgtt tattgcagct tataatggtt acaaataaag
     3721 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3781 gtccaaactc atcaatgtat cttatcatgt ctggatcggg aattaattcg gcgcagcacc
     3841 atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa
     3901 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3961 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4021 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4081 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4141 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4201 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctgt taacagcttg
     4261 gctcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4321 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4381 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4441 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4501 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4561 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4621 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtt
//