LOCUS       Exported                4955 bp ds-DNA     circular SYN 27-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4955)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Oct 27, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4955
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(336..1196)
                     /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"
     CDS             complement(339..998)
                     /label=AmpR
     primer_bind     complement(386..405)
                     /label=AmpStop
     primer_bind     1132..1151
                     /label=AmpStart
     promoter        complement(1197..1301)
                     /gene="bla"
                     /label=AmpR promoter
     gap             1238..1266
                     /label=amp prom|Unknown
     rep_origin      complement(1346..1801)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(1478..1784)
                     /direction=LEFT
                     /label=F1 ori
     CDS             complement(1803..1871)
                     /label=LacZ alpha
     primer_bind     1891..1910
                     /label=pUCM13-52
     primer_bind     1923..1939
                     /label=M13-40
     primer_bind     1942..1959
                     /label=M13-fwd
     primer_bind     1942..1958
                     /label=M13
     primer_bind     1943..1959
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1968..1988
                     /label=T7
     primer_bind     1969..1988
                     /label=T7
     promoter        1969..1987
                     /label=T7_promoter Promoter|promoter
     primer_bind     1969..1987
                     /label=Short-T7
     misc_feature    2012..2390
                     /label=P_TEF1
     primer_bind     2012..2030
                     /label=O5447
     primer_bind     2012..2030
                     /label=FRO4618
     primer_bind     2012..2030
                     /label=o5442
     primer_bind     2012..2030
                     /label=o5460
     primer_bind     2012..2030
                     /label=o5455
     primer_bind     2042..2061
                     /label=FRO351
     misc_feature    2046..2385
                     /label=pTEF-short
     gene            2047..3166
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        2047..2390
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     misc_feature    2289..2296
                     /label=TATAbox
     primer_bind     2361..2390
                     /label=FRO4538
     primer_bind     complement(2361..2390)
                     /label=FRO4540
     primer_bind     2371..2411
                     /label=FRP373 FWD
     misc_feature    2391..2963
                     /label=Resistance Marker
     misc_feature    2391..2963
                     /label=NatMX
     CDS             2391..2963
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     primer_bind     complement(2478..2497)
                     /label=FRO1851
     misc_feature    2964..3200
                     /label=term_TEF1
     misc_feature    2964..3164
                     /label=TermTef-Short
     primer_bind     2964..2993
                     /label=FRO4541
     primer_bind     complement(2964..2993)
                     /label=FRO4539
     terminator      2969..3166
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3178..3201)
                     /label=FRO4760
     primer_bind     complement(3199..3222)
                     /label=FRO4299
     primer_bind     3202..3246
                     /label=FRO4300
     primer_bind     3217..3250
                     /label=o5450
     primer_bind     3222..3250
                     /label=o5456
     misc_feature    3223..3947
                     /label=P7Sul.1
     primer_bind     3223..3250
                     /label=FRO4507
     primer_bind     3223..3250
                     /label=o5451
     primer_bind     3223..3250
                     /label=o5462
     primer_bind     complement(3228..3249)
                     /label=FRO4368
     primer_bind     complement(3235..3254)
                     /label=FRO4445
     primer_bind     complement(3301..3346)
                     /label=o5259
     misc_feature    3321..3336
                     /label=minSulA
     primer_bind     3327..3383
                     /label=o5260
     misc_feature    3336..3347
                     /label=RAP1binding
     misc_feature    3348..3363
                     /label=minSulA
     misc_feature    3363..3373
                     /label=GCR1binding
     primer_bind     3384..3403
                     /label=O5278
     misc_feature    3385..3394
                     /label=grf2_binding
     primer_bind     complement(3410..3468)
                     /label=FRO4981
     primer_bind     complement(3410..3428)
                     /label=FRO5013
     misc_feature    3415..3425
                     /label=grf2_binding
     misc_feature    3429..3444
                     /label=minSulA
     misc_feature    3444..3455
                     /label=RAP1binding
     misc_feature    3456..3471
                     /label=minSulA
     primer_bind     3469..3534
                     /label=FRO4980
     primer_bind     3471..3490
                     /label=FRO4293
     misc_feature    3471..3481
                     /label=GCR1binding
     primer_bind     complement(3482..3519)
                     /label=FRO5169
     misc_feature    3494..3509
                     /label=minSulA
     misc_feature    3509..3519
                     /label=GCR1binding
     primer_bind     3518..3547
                     /label=FRO5023
     primer_bind     3518..3534
                     /label=FRO5014
     primer_bind     3732..3755
                     /label=o5203
     primer_bind     complement(3732..3755)
                     /label=o5202
     primer_bind     complement(3782..3825)
                     /label=o5261
     misc_feature    3797..3817
                     /label=sulA
     misc_feature    3802..3819
                     /label=sulA.18
     misc_feature    3802..3817
                     /label=minSulA
     primer_bind     3805..3864
                     /label=o5262
     misc_feature    3818..3825
                     /label=TATAbox
     misc_feature    3826..3844
                     /label=sulA
     misc_feature    3826..3843
                     /label=sulA.18
     misc_feature    3826..3841
                     /label=minSulA
     primer_bind     3842..3862
                     /label=FRO4729
     primer_bind     complement(3909..3937)
                     /label=FRO5163
     primer_bind     3909..3927
                     /label=FRO0923
     primer_bind     complement(3916..3938)
                     /label=o5454
     primer_bind     complement(3916..3938)
                     /label=o5448
     primer_bind     complement(3916..3938)
                     /label=o5443
     primer_bind     complement(3916..3938)
                     /label=o5461
     primer_bind     3927..3955
                     /label=FRO4431
     primer_bind     complement(3927..3952)
                     /label=FRO5079
     primer_bind     complement(3932..4022)
                     /label=FRO4297
     misc_feature    3938..3952
                     /label=good kozak
     primer_bind     3950..4015
                     /label=FRO4855
     primer_bind     3955..3991
                     /label=FRO4855
     gap             3956..4027
                     /label=FLAG TAG|Unknown
     CDS             3956..4027
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(3965..3994)
                     /label=FRO4412
     primer_bind     complement(3965..3984)
                     /label=FRO4408
     primer_bind     complement(3989..4018)
                     /label=FRO4412
     primer_bind     complement(3989..4008)
                     /label=FRO4408
     primer_bind     complement(3996..4051)
                     /label=FRO4856
     primer_bind     4002..4092
                     /label=FRO4298
     primer_bind     complement(4013..4028)
                     /label=FRO4408
     primer_bind     complement(4013..4028)
                     /label=FRO4412
     gap             4028..4051
                     /label=8G linker|Unknown
     primer_bind     complement(4052..4075)
                     /label=FRO4532
     primer_bind     4052..4072
                     /label=S4 primer site|primer_bind
     primer_bind     complement(4073..4097)
                     /label=FRO4432
     primer_bind     complement(4089..4108)
                     /label=T3
     primer_bind     complement(4090..4109)
                     /label=FRO0114
     promoter        complement(4091..4109)
                     /label=T3 promoter
     primer_bind     complement(4091..4107)
                     /label=T3
     primer_bind     complement(4126..4146)
                     /label=M13-rev
     primer_bind     complement(4130..4146)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4130..4146)
                     /label=M13r
     primer_bind     4144..4168
                     /label=FRO2169
     primer_bind     complement(4144..4163)
                     /label=LacOp-for
     misc_binding    complement(4152..4174)
                     /label=LacO
     misc_feature    complement(4152..4172)
                     /label=lacop
     misc_feature    complement(4154..4172)
                     /label=lacop
     protein_bind    4154..4170
                     /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(4178..4208)
                     /label=lac promoter
     primer_bind     complement(4178..4197)
                     /label=pUCM13-rev-157
     protein_bind    4223..4244
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(join(4514..4955,1..241))
                     /direction=LEFT
                     /label=ColE1 origin
     rep_origin      complement(join(4532..4955,1..165))
                     /direction=LEFT
                     /label=ori|rep_origin
ORIGIN
        1 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
       61 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      121 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      181 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      241 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      301 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
      361 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
      421 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
      481 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
      541 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
      601 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
      661 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
      721 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
      781 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
      841 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
      901 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
      961 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1021 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1081 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1141 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1201 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     1261 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     1321 ccaccaatac ccgactctat atgctaaatt gtaagcgtta atattttgtt aaaattcgcg
     1381 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     1441 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     1501 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     1561 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     1621 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     1681 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     1741 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     1801 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     1861 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     1921 gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca
     1981 ctatagggcg aattggagct cgatcctcga gagcttgcct tgtccccgcc gggtcacccg
     2041 gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg
     2101 gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca
     2161 tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcggacc
     2221 tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg
     2281 tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt
     2341 aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc atgggtacca
     2401 ctcttgacga cacggcttac cggtaccgca ccagtgtccc gggggacgcc gaggccatcg
     2461 aggcactgga tgggtccttc accaccgaca ccgtcttccg cgtcaccgcc accggggacg
     2521 gcttcaccct gcgggaggtg ccggtggacc cgcccctgac caaggtgttc cccgacgacg
     2581 aatcggacga cgaatcggac gacggggagg acggcgaccc ggactcccgg acgttcgtcg
     2641 cgtacgggga cgacggcgac ctggcgggct tcgtggtcgt ctcgtactcc ggctggaacc
     2701 gccggctgac cgtcgaggac atcgaggtcg ccccggagca ccgggggcac ggggtcgggc
     2761 gcgcgttgat ggggctcgcg acggagttcg cccgcgagcg gggcgccggg cacctctggc
     2821 tggaggtcac caacgtcaac gcaccggcga tccacgcgta ccggcggatg gggttcaccc
     2881 tctgcggcct ggacaccgcc ctgtacgacg gcaccgcctc ggacggcgag caggcgctct
     2941 acatgagcat gccctgcccc taatcagtac tgacaataaa aagattcttg ttttcaagaa
     3001 cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt
     3061 gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa
     3121 agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata
     3181 ctaacgccgc catccagtgt ttaaacgaac tagtgcggcc gccagttcga gtttatcatt
     3241 atcaatactg ccatttcaaa gaatacgtaa ataattaata gtagtgattt tcctaacttt
     3301 atttagtcaa aaaattagcc ctgtacatcc atacagtgtc tgggtgactg tacatccata
     3361 cagcatccac taattttaat tctgctgtaa cccgtacatg cccaaaatag ggggcgggtt
     3421 acacagaact gtacatccat acagtgtctg ggtgactgta catccataca gcatccacta
     3481 aatataatgg agcctgtaca tccatacagc atccactaaa aaaaagaatc ccagcaccaa
     3541 aatattgttt tcttcaccaa ccatcagttc ataggtccat tctcttagcg caactacaga
     3601 gaacaggggc acaaacaggc aaaaaacggg cacaacctca atggagtgat gcaacctgcc
     3661 tggagtaaat gatgacacaa ggcaattgac ccacgcatgt atctatctca ttttcttaca
     3721 ccttctatta ccttctgctc tctctgattt ggaaaaagct gaaaaaaaag gttgaaacca
     3781 gttccctgaa attattcccc tctgtacatc catacagtat aaatactgta catccataca
     3841 gtaattctgt aaatctattt cttaaacttc ttaaattcta cttttatagt tagtcttttt
     3901 tttagtttta aaacaccaag aacttagttt cgaataaaca cacataaaca aaatggatta
     3961 taaagatgac gatgacaagg attataaaga tgacgatgac aaggattata aagatgacga
     4021 tgacaaggga ggtggtggtg gtggaggcgg tccgacagag aattcatcga tgggtaccca
     4081 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     4141 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4201 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4261 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4321 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4381 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4441 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4501 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4561 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4621 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4681 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4741 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4801 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4861 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4921 gcggtgctac agagttcttg aagtggtggc ctaac
//
