LOCUS       sequence-486018-        4506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBAD-SD8-GIII Neg; AP Neg Circuit for AraC phage-assisted evolution.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4506)
  TITLE     Phage-Assisted Evolution of Allosteric Protein Switches
REFERENCE   2  (bases 1 to 4506)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: PACE_AP_Neg_pBADProm
FEATURES             Location/Qualifiers
     source          1..4506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     479..498
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     732..749
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    764..904
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(830..849)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     990..1012
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1006..1197)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    1858..1877
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        1884..2168
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2091..2110
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2288..2305
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      complement(3386..3432)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3483..4274,4275..4343))
                     /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     4106..4125
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4344..4448)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(4496..4506,1..578)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
       61 gtttgccgga tcaagagcta ccacctcttt ttccgaaggt aactggcttc agcagagcgc
      121 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
      181 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
      241 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
      301 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
      361 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
      421 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
      481 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
      541 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
      601 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
      661 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
      721 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
      781 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
      841 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
      901 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
      961 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     1021 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     1081 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     1141 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     1201 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     1261 gatgctcacg atacgggtta ctgatgatga acagaggaca tccggtaagg ccaatagggg
     1321 ttcataaggt acggcccgta ctgaacccgg gtaggctgct tgagccagtg agcgattgct
     1381 ggcctagatg aatgactgtc cacgacagaa cccggcttat cggtcagttt cacctgattt
     1441 acgtaaaaac ccgcttcggc gggtttttgc ttttggaggg gcagaaagat gaatgactgt
     1501 ccacgacgct atacccaaaa gaaagcggct tatcggtcag tttcacctgg tttacgtaaa
     1561 aacccgcttc ggcgggtttt tgcttttgga ggggcagaaa gatgaatgac tgtccacgac
     1621 actataccca aaagaaagcg gcttatcggt cagtttcacc tgttttacgt aaaaacccgc
     1681 ttcggcgggt ttttactttt ggaggggcag aaagatgaat gactgtccac gacactatac
     1741 ccaaaagaaa gcggcttatc ggtcagtttt acctgatgta cgtaataaac cgttccggcg
     1801 ggtttatgcc tttaattaag gagttttgca ggtgccttgg aacacctgct tttcgctgaa
     1861 ttcgcggccg cttctagact agaaagaaac caattgtcca tattgcatca gacattgccg
     1921 tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag
     1981 cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa
     2041 tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc
     2101 atttttatcc ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt
     2161 ttctccatga ccgaattcaa aagatctttt aagaaggaga tatacaaagg aggaaaaaaa
     2221 aatgaaaaaa ttattattcg caattccttt agttgttcct ttctattctc actccgctga
     2281 aactgttcat caccatcacc atcacgctga aactgttgaa agttgtttag caaaacccca
     2341 tacagaaaat tcatttacta acgtctggaa agacgacaaa actttagatc gttacgctaa
     2401 ctatgagggc tgtctgtgga atgctacagg cgttgtagtt tgtactggtg acgaaactca
     2461 gtgttacggt acatgggttc ctattgggct tgctatccct gaaaatgagg gtggtggctc
     2521 tgagggtggc ggttctgagg gtggcggttc tgagggtggc ggtactaaac ctcctgagta
     2581 cggtgataca cctattccgg gctatactta tatcaaccct ctcgacggca cttatccgcc
     2641 tggtactgag caaaaccccg ctaatcctaa tccttctctt gaggagtctc agcctcttaa
     2701 tactttcatg tttcagaata ataggttccg aaataggcag ggggcattaa ctgtttatac
     2761 gggcactgtt actcaaggca ctgaccccgt taaaacttat taccagtaca ctcctgtatc
     2821 atcaaaagcc atgtatgacg cttactggaa cggtaaattc agagactgcg ctttccattc
     2881 tggctttaat gaggatccat tcgtttgtga atatcaaggc caatcgtctg acctgcctca
     2941 acctcctgtc aatgctggcg gcggctctgg tggtggttct ggtggcggct ctgagggtgg
     3001 tggctctgag ggtggcggtt ctgagggtgg cggctctgag ggaggcggtt ccggtggtgg
     3061 ctcttcccaa atggctcaag tcggtgacgg tgataattca cctttaatga ataatttccg
     3121 tcaatattta ccttccctcc ctcaatcggt tgaatgtcgc ccttttgtct ttggcgctgg
     3181 taaaccttac gagttcagta tcgactgcga taagatcaac ctgttccgcg gtgtctttgc
     3241 gtttctttta tatgttgcca cctttatgta tgtattttct acgtttgcta acatactgcg
     3301 taataaggag tcttaaaaat ataatgaccc tcttgataac ccaagagggc attttttaat
     3361 gcccatggcg tagctgtgct ctagaataaa acgaaaggcc cagtctttcg actgagcctt
     3421 tcgttttatt tgaccggaaa tcagaattgg ttaattggtt gtaacactgg cagagcatta
     3481 cgttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3541 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3601 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3661 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3721 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3781 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3841 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3901 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3961 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4021 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4081 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4141 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4201 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4261 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4321 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4381 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4441 gttccgcgat taattgcgtt gcgctcactg cccgctttcc aggccaaata ggccgttgag
     4501 atcctt
//
