LOCUS       Exported                5574 bp ds-DNA     circular SYN 26-SEP-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5574)
  AUTHORS   Vojislav
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Sep 30, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5574
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     gene            complement(249..1824)
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     terminator      249..446
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(452..1480)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(1481..1824)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     rep_origin      complement(1941..2396)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2078..2099)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2290..2309
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2373..2626)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="SIPSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRL
                     AAHPPFASWRNSEEARTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2522..2544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2536..2553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2537..2553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2563..2582
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2563..2581
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2590..2626
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2606..2622
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2607..2623
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2632..2978
                     /label=3' YOR1 UTR
     misc_feature    2979..3269
                     /label=5' YOR1 UTR
     primer_bind     complement(3246..3296)
                     /label=Fragment 2.REV
     primer_bind     complement(3271..3287)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(3275..3367)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELV"
     misc_feature    3275..3311
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     complement(3275..3287)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3324..3344)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3324..3342)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3363..3379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3363..3379)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3376..3398)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3387..3403
                     /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(3411..3441)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3456..3477
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3594..3611)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3765..4353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3845..3864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4524..5384)
                     /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     5147..5166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5385..5489)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataatca tcgatgaatt cgagctcgtt ttcgacactg gatggcggcg ttagtatcga
      241 atcgacagca gtatagcgac cagcattcac atacgattga cgcatgatat tactttctgc
      301 gcacttaact tcgcatctgg gcagatgatg tcgaggcgaa aaaaaatata aatcacgcta
      361 acatttgatt aaaatagaac aactacaata taaaaaaact atacaaatga caagttcttg
      421 aaaacaagaa tctttttatt gtcagtactg attattcctt tgccctcgga cgagtgctgg
      481 ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag acggccgcgc
      541 ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg attgcgtcgc
      601 atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc tgatagagtt
      661 ggtcaagacc aatgcggagc atatacgccc ggagccgcgg cgatcctgca agctccggat
      721 gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc ctccagaaga
      781 agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag tcaatgaccg
      841 ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg tgcacgaggt
      901 gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc tgcgcgacgg
      961 acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga tcagcaatcg
     1021 cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg aatgggccga
     1081 acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catggcctcc gcgaccggct
     1141 gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc tgtgcacggc
     1201 gggagatgca ataggtcagg ctctcgctga attccccaat gtcaagcact tccggaatcg
     1261 ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa ccatcggcgc
     1321 agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa gcacgagatt
     1381 cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg atcagaaact
     1441 tctcgacaga cgtcgcggtg agttcaggct ttttacccat ggttgtttat gttcggatgt
     1501 gatgtgagaa ctgtatccta gcaagatttt aaaaggaagt atatgaaaga agaacctcag
     1561 tggcaaatcc taacctttta tatttctcta caggggcgcg gcgtggggac aattcaacgc
     1621 gtctgtgagg ggagcgtttc cctgctcgca ggtccgcagc gaggagccgt aatttttgct
     1681 tcgcgccgtg cggccatcaa aatgtatgga tgcaaatgat tatacatggg gatgtatggg
     1741 ctaaatgtac gggcgacagt cacatcatgc ccctgagctg cgcacgtcaa gactgtcaag
     1801 gagggtattc tgggcctcca tgtcgctggc cgggtgaccc ggcggggacg aggcaagcta
     1861 aacagatctg gcgcgcctta attaacccta tgcggtgtga aataccgcac agatgcgtaa
     1921 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     1981 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2041 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2101 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2161 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2221 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2281 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     2341 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     2401 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     2461 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     2521 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag
     2581 ggcgaattgg gtaccgggcc ccccctcgag gtcgacggta tcgataagct taaaacttag
     2641 aagtgaaaat cctaaaaccg aaggatattt caaatacgta aaagaagtga aagataaaat
     2701 aaagcctaaa taaggaagaa aagaagggat aacatttttt tttgttactt tttgcttatt
     2761 cctcacctaa acagaaggaa aaagccattc ttgtttaaag agtattttta aagcgttcat
     2821 gaaaagtcac agtccaagga gtatttcaaa ttgtcacttg aagtgaaaac accccagtgc
     2881 ttctcgacat cagaggtacc agaggtgttt ggcttccaat cttcatcaaa ggcttcaaaa
     2941 acaataacgt taacacccca agctctcatg gaacagatat ctgctgtccc ttaaatgatt
     3001 accatacata tatatattta ttacacatct gtcagaggta gctagcgaag gtgtcactga
     3061 aatatttttt gttccagtta gtataaatac ggaggtagaa cagctctccg cgtgtatatc
     3121 tttttttgcg ctatacaaga acaggaagaa cgcatttcca tacctttttc tccttacagg
     3181 tgccctctga gtagtgtcac gaacgaggaa aaagattaat attactgttt ttatattcaa
     3241 aaagagtaaa gccgttgcta tatacgaatg gatccactag ttctagagcg gccgccaccg
     3301 cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     3361 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3421 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3481 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3541 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3601 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3661 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3721 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3781 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3841 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3901 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3961 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4021 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4081 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4141 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4201 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4261 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4321 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4381 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4441 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4501 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4561 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4621 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4681 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4741 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4801 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4861 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4921 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4981 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5041 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5101 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5161 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5221 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5281 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5341 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5401 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5461 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     5521 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gagg
//
