LOCUS       Exported                5198 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pInt
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5198)
  AUTHORS   Groth AC, Olivares EC, Thyagarajan B, Calos MP
  TITLE     A phage integrase directs efficient site-specific integration in 
            human cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2000 May 23. 97(11):5995-6000.
  PUBMED    10801973
REFERENCE   2  (bases 1 to 5198)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5198
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(16..1833)
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     primer_bind     complement(1897..1917)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1897..1915)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1936..1952)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1936..1952)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1949..1971)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1960..1976
                     /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(1984..2014)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2029..2050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2662..3206
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     3107..3126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3323..3340
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3790..3809)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3801..4616
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3874..3893)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gatccgggtg tctcgctacg ccgctacgtc ttccgtgccg tcctgggcgt cgtcttcgtc
       61 gtcgtcggtc ggcggcttcg cccacgtgat cgaagcgcgc ttctcgatgg gcgttccctg
      121 ccccctgccc gtagtcgact tcgtgacaac gatcttgtct acgaagagcc cgacgaacac
      181 gcgcttgtcg tctactgacg cgcgccccca ccacgactta gggccggtcg ggtcagcgtc
      241 ggcgtcttcg gggaaccatt ggtcaagggg aagcttcggg gcttcggcgg cttcaagttc
      301 ggcaagccgc tcttccgccc cttgctgccg gagcgtcagc gctgcctgtt gcttccggaa
      361 gtgcttcctg ccaacgggtc cgtcgtacgc gcctgccgcg cggtcttcgt acagctcttc
      421 aagggcgttc agggcgtcgg cgcgctccgc aacaaggttc gcccgttcgc cgctcttctc
      481 aggcgcctca gtgagcttgc cgaagcgtcg ggcggcttcc cacagaagcg ccaacgtctc
      541 ttcgtcgcct tcggcgtgcc tgatcttgtt gaagatgcgt tccgcaacga acttgtcgag
      601 tgccgccatg ctgacgttgc acgtgccttc gtgctgccca ggtgcggacg ggtcgaccac
      661 cttccggcga cggcagcggt aagagtcctt gatcgattct tccccgcgct tcgaagtcat
      721 gacggcgcca cactcgcagt acagcttgtc catggcggac agaatggctt gcccccggga
      781 aagccccttg ccgcgccccc tgccgtccaa ccacgcctga agctcatacc actcagcggg
      841 ctcgatgatc ggtccgcaat caagctcgac cggccggagc gtgatcgggt cgcgctgaat
      901 gcggtaaccc tcaatcttcg tggtcggcgt gccgtccggc ttcttcttgt agatcacctc
      961 agcggcgaag cccgcaatac gcgggtcccg aaggattcgc ataacggttg ccgggtccca
     1021 ggcgcttgaa gcggtcttct tcccaatcgt ctcgccccgg gtcggcacgg cgtcagcgtc
     1081 catgcgctta caaagccccg tgatgctgcc cgggtgaatg gcggcttgac tgcccggctt
     1141 gaagggaagg tgtttgtgcg tcttgatctc acgccaccac caccggatta cgtcgggctc
     1201 gaactcgaag ggtccggtaa ggggagtggt cgagtgcgca agcttgttga tgacgacatt
     1261 gaccattcgg ccgttgcgcg tgatctcctt cgtctccgaa acaagctcga agccgtaagg
     1321 cgccttcccg ccgacgtacc cgcccaattc gcgctgaagg ttcttcgtgt cgagaatctt
     1381 cgccgacttc agcgaagatt ctttgtgcga cgcgtcgagc cgcataatca ggtgaatcag
     1441 gtccatgacg tttccctgcc ggaagacgcc ttcctgagtg gaaacaatcg tcacgcccag
     1501 ggcgagcaat tccgagacaa tcggaatcgc gtccatgacc ttcaggcgcg agaagcgcga
     1561 cacgtcatag acaatgatca tgttgagccg cccggcgcgg cattcgttca ggatgcgttc
     1621 gaactccggg cgctccgccg tcccgaacgc cgacgtgccc ggcgcttcgc tgaaatgccc
     1681 gacgaacctg aaccggcccc cgtcgcgctc gacttcgcgc tgaaggtcgg ccgccttgtc
     1741 ttcgttggcg ctacgctgtg tcgctgggct tgctgcgctc gaattctcgc gctcgcgcga
     1801 ctgacggtcg taagcacccg cgtacgtgtc caccccggtc acaacccctt gtgtcatgtc
     1861 ggcgacccta cgactagtga gctccagctt ttgttccctt tagtgagggt taattgcgcg
     1921 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1981 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2041 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2101 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2161 cgctcggcca aaaaggcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     2221 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2281 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2341 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2401 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2461 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2521 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2581 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2641 ccccttaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga
     2701 aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt
     2761 gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc
     2821 ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca
     2881 gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc
     2941 agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc
     3001 cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg
     3061 gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccagggga aacgcctggt
     3121 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct
     3181 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt
     3241 aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg
     3301 ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta
     3361 tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca
     3421 ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag cgctgaggtc
     3481 tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca tcatccagcc
     3541 agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag ttggtgattt
     3601 tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct
     3661 tcaactcagc aaaagttcga tttattcaac aaagccacgt tgtgtctcaa aatctctgat
     3721 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     3781 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc tcgaggccgc
     3841 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     3901 ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc
     3961 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     4021 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     4081 catggttact caccactgcg atccccggga aaacagcatt ccaggtatta gaagaatatc
     4141 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     4201 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     4261 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     4321 ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg gattcagtcg
     4381 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     4441 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     4501 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     4561 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     4621 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcggct
     4681 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa
     4741 cgcagaccgt tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa
     4801 agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcctg
     4861 gtatgagtca gcaacacctt cttcacgagg cagacctcag cgctcaaaga tgcaggggta
     4921 aaagctaacc gcatctttac cgacaaggca tccggcagtt caacagatcg ggaagggctg
     4981 gatttgctga ggatgaaggt ggaggaaggt gatgtcattc tggtgaagaa gctcgaccgt
     5041 cttggccgcg acaccgccga catgatccaa ctgataaaag agtttgatgc tcagggtgta
     5101 gcggttcggt ttattgacga cgggatcagt accgacggtg atatggggca aatggtggtc
     5161 accatcctgt cggctgtggc acaggctgaa cgccggag
//
