LOCUS       sequence-47839-e        5069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5069)
  AUTHORS   Watabe-Uchida M, Zhu L, Ogawa SK, Vamanrao A, Uchida N
  TITLE     Whole-brain mapping of direct inputs to midbrain dopamine neurons.
  JOURNAL   Neuron. 2012 Jun 7;74(5):858-73.
  PUBMED    22681690
REFERENCE   2  (bases 1 to 5069)
  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"; journalName: "Neuron."; 
            date: "2012-06-7"; volume: "74(5)"; pages: "858-73"
COMMENT     Sequence Label: pCA-FLEX
FEATURES             Location/Qualifiers
     source          1..5069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(27..455)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(137..158)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     349..368
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     582..604
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     596..613
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     597..613
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     674..690
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     674..690
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     enhancer        769..1072
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1075..1350
                     /label=chicken beta-actin promoter
     intron          1351..2359
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2283..2305
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     polyA_signal    2709..2843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2746..2765)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2800..2819
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2882..2902)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2882..2900)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2921..2937)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2921..2937)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2934..2956)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2945..2961
                     /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(join(2969..2975,2976..2993,2994..2999))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3014..3035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3152..3169)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3323..3911)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3403..3422)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4082..4873,4874..4942))
                     /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     4705..4724
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4943..5047)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
       61 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
      121 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
      181 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
      241 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
      301 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
      361 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
      421 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
      481 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
      541 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
      601 aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg agctccaccg
      661 cggtggcggc cgctctagaa ctagtggatc ccccgggctg caggaatcta gttattaata
      721 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      781 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      841 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggacta
      901 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      961 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     1021 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgggtcgagg
     1081 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
     1141 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggcgcgcg
     1201 ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca
     1261 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
     1321 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgttgccttc gccccgtgcc
     1381 ccgctccgcg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
     1441 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
     1501 ctcgtttctt ttctgtggct gcgtgaaagc cttaaagggc tccgggaggg ccctttgtgc
     1561 gggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
     1621 cccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
     1681 gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg gctgcgaggg
     1741 gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcggc
     1801 ggtcgggctg taaccccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt
     1861 cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc
     1921 ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc tcgggggagg
     1981 ggcgcggcgg ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct
     2041 tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct ggcggagccg
     2101 aaatctggga ggcgccgccg caccccctct agcgggcgcg ggcgaagcgg tgcggcgccg
     2161 gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgcggt ccccttctcc
     2221 atctccagcc tcggggctgc cgcaggggga cggctgcctt cgggggggac ggggcagggc
     2281 ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc
     2341 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt gttgtgctgt ctcatcattt
     2401 tggcaaagaa ttgatttcga tacgtacggt tgaattcatt acctcgtata ggatacttta
     2461 tacgaagtta tgcagaatgg tagctggatt gtaagtacta ttagcaatat gaaacctctt
     2521 aataacctcg tatagcatac attatacgaa gtaatagatc taagcttgat atcgtcgaca
     2581 taacttcgta taaagtatcc tatacgaggt aatttgcctt aacccagaaa ttatcagtac
     2641 tattctttag aatggtgcaa agaattactt cgtataatgt atgctatacg aggttatcgc
     2701 atgctagcaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2761 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2821 atgtatctta tcatgtctgg atctcgacct cgaggggggg cccggtaccc agcttttgtt
     2881 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     2941 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3001 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3061 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3121 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3181 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3241 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3301 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3361 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3421 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3481 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3541 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3601 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3661 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3721 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     3781 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3841 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3901 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3961 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4021 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4081 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4141 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4201 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4261 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4321 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4381 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4441 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4501 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4561 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4621 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4681 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4741 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4801 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4861 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4921 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4981 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5041 ttccgcgcac atttccccga aaagtgcca
//
