LOCUS       Exported                5504 bp DNA     circular SYN 20-JUN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5504)
  AUTHORS   Vahid Asimi
  TITLE     Direct Submission
  JOURNAL   Exported Jun 21, 2022 from SnapGene 6.0.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             216..270
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDP"
     primer_bind     231..297
                     /label=Fragment 1.FOR
     misc_feature    233..270
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     misc_feature    271..1070
                     /label=5' Homology Region TRIM28
     primer_bind     1053..1099
                     /label=Fragment 2.FOR
     primer_bind     complement(1057..1092)
                     /label=Fragment 1.REV
     misc_feature    1057..1072
                     /label=sgRNA1
     misc_feature    1071..1072
                     /label=to correct frame
     misc_feature    1073..1789
                     /label=mCerulean
     misc_feature    1790..1797
                     /label=GS LInker
     CDS             1856..1882
                     /label=HA
     CDS             1886..1912
                     /label=HA
     CDS             1922..2242
                     /label=FKBP-V
     misc_feature    2243..2287
                     /label=glycinlinker
     primer_bind     2268..2301
                     /label=Fragment 3.FOR
     primer_bind     complement(2271..2307)
                     /label=Fragment 2.REV
     misc_feature    2288..2294
                     /label=sgRNA1
     misc_feature    2288
                     /label=to correct frame
     misc_feature    2289..3088
                     /label=3' Homology Region TRIM28
     primer_bind     complement(2439..2451)
                     /label=Fragment 2.REV
     primer_bind     complement(3072..3128)
                     /label=Fragment 3.REV
     CDS             3089..3357
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="VPSSNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTD
                     RPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDAA"
     misc_feature    3089..3107
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3108..3124)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3598..3702
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3703..4563
                     /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      4734..5322
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gtcgactcta gaggatcccc ccgagtttaa caataagaaa gtttgggtta
      301 agaccgcctc cacgttcggc gggaacttcc tggcaaagac cccctcaccc cctccacctg
      361 ggctccggcg ctttgctctg ccgtggcccc gcccaccctg tattgtcccg cccctcttcc
      421 ccgactatag gccaggctgc cgctgcagac gtcgccacgc ctgctgagga ggcggggcct
      481 aaggttcggc tctgcaggac ctgcgcgctc aagcacgcgg gttgccagcc tgggtgcacc
      541 gtgcggtggg catgggagac gtcaagcacg cactctgcgg gtgaattcct caccagcagc
      601 attgctactc tggatctaga ggggcggcag cgagtggaag ggagggccct ggcgcgtgcg
      661 taatctgcgc cccgccccca ggctgcggtg cccaacgcgc gtgcgcggcg tcgtcggcgc
      721 cagatatttc tgtcccgcct ccaggccgcg gctctttctg cgagcggcgc gcggtcaggc
      781 ggttgttcgc aagtcttgcg gcgcgagtgt ctcagaagcg agggaggagg cggcgctggt
      841 ggcggcgccg tcgtctgtac agaggcccgc agccagcggc ttggccgcat ctcgacagcg
      901 cctggggcgc ggcgggcgtc ggcccaggag acgcgtggcg gcgctcggcc tcgcggcatc
      961 ggcggctgcc tggccgttgg cggcgagcgc acttgcgcct gcgcagcggg ctccgtgccc
     1021 ctcctcccct gggcggcccc cccacccccc cggcggcgtg tgaatggcgg ccatggtgag
     1081 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1141 aaacggccac aggttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1201 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1261 caccctgacc tggggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     1321 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgtaccatct tcttcaagga
     1381 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1441 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1501 gtacaacgcc atcagcgaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     1561 ggcccacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1621 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1681 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1741 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gaagcggagc
     1801 tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg gaccttaccc
     1861 ctacgacgtg cccgactacg ccggctatcc gtatgatgtc ccggactatg caggaagcgg
     1921 aggagtgcag gtggaaacca tctccccagg agacgggcgc accttcccca agcgcggcca
     1981 gacctgcgtg gtgcactaca ccgggatgct tgaagatgga aagaaagttg attcctcccg
     2041 ggacagaaac aagcccttta agtttatgct aggcaagcag gaggtgatcc gaggctggga
     2101 agaaggggtt gcccagatga gtgtgggtca gagagccaaa ctgactatat ctccagatta
     2161 tgcctatggt gccactgggc acccaggcat catcccacca catgccactc tcgtcttcga
     2221 tgtggagctt ctaaaactgg aaggtggcgg tggctcgggc ggtggtgggt cgggtggcgg
     2281 cggatctgcc tcggcggcag cgactgcagc ggcctcggcc gcgacggccg cctcggcggc
     2341 ctctggtagc ccagggtcgg gcgagggctc ggcgggcggt gagaagcgtc cggctgcttc
     2401 ctcagccgcg gcggcctctg cagccgcgtc gtcccctgcg gggggcggtg gcgaggcgca
     2461 ggagcttctg gagcactgcg gcgtgtgtcg cgagcgcctg cggcccgagc gggatcctcg
     2521 gctgctgccc tgtctacatt cggcctgcag tgcctgcctg ggccccgcta cacccgccgc
     2581 agcgaataat tcgggggatg gcggctcggc gggcgacggc gctagtgagt actgggtggc
     2641 caggtgcccc tccccctcct cgcagcccgt gctcgggact gcgcctgtgc gagagtatgg
     2701 gggcccgggt agggttaagt aggcctgctg gtttaagagg gggcggggaa cgggtcctgg
     2761 cctctgccaa tgcccgttac caggtctgga caccgaggtg cagaatgtga tgggagatgt
     2821 caagaaatca gagggtgtgc atgcattccc atttttatct tggcagtggt ggattgtcca
     2881 gtgtgcaaac agcagtgcta ctccaaagac atcgtggaga attattttat gcgtgatagt
     2941 ggcagtaagg cctcttctga ttcccaggat gctaaccagg tatgtcctgt gggaaatctt
     3001 ggggaagagg gaagcgtgac cttttccaga gcctctcacc atcttcacta gtttggattg
     3061 tatccttatg cggtctcagt gtctgggagg gtaccgagct cgaattcact ggccgtcgtt
     3121 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     3181 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     3241 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     3301 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3361 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3421 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3481 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3541 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     3601 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3661 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3721 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3781 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3841 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3901 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     3961 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4021 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4081 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4141 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4201 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4261 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4321 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4381 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4441 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4501 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4561 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4621 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4681 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4741 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4801 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4861 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4921 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4981 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5041 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5101 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5161 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5221 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5281 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5341 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5401 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     5461 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaga
//
